Load libraries (packages)

library("respR") ## respirometry/slope analysis
library("tidyverse") ## data manipulation

Set working directory

setwd("[PATH TO DIRECTORY]")

System1 - Dell

Importing data from firesting for resting

preexperiment_date <- "16 May 2023 09 49AM/All"
postexperiment_date <- "16 May 2023 03 19PM/All"

##--- last fish run in trial ---##
experiment_date <- "16 May 2023 11 29AM/Oxygen"
experiment_date2 <- "16 May 2023 11 29AM/All"

firesting <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",experiment_date,"data raw/Firesting.txt"), 
    delim = "\t", escape_double = FALSE, 
    col_types = cols(`Time (HH:MM:SS)` = col_time(format = "%H:%M:%S"), 
        `Time (s)` = col_number(), Ch1...5 = col_number(), 
        Ch2...6 = col_number(), Ch3...7 = col_number(), 
        Ch4...8 = col_number()), trim_ws = TRUE, 
    skip = 19)

Cycle_1 <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",experiment_date2,"slopes/Cycle_1.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 

Cycle_last <-read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",experiment_date2,"slopes/Cycle_21.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 

System2 - Asus

Importing data from firesting for resting

preexperiment_date_asus <- "16 May 2023 10 21AM/All"
postexperiment_date_asus <- "16 May 2023 03 49PM/All"

##--- last fish run in trial ---##
experiment_date_asus <- "16 May 2023 12 38PM/Oxygen"
experiment_date2_asus <- "16 May 2023 12 38PM/All"

firesting_asus <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",experiment_date_asus,"data raw/Firesting.txt"), 
    delim = "\t", escape_double = FALSE, 
    col_types = cols(`Time (HH:MM:SS)` = col_time(format = "%H:%M:%S"), 
        `Time (s)` = col_number(), Ch1...5 = col_number(), 
        Ch2...6 = col_number(), Ch3...7 = col_number(), 
        Ch4...8 = col_number()), trim_ws = TRUE, 
    skip = 19)

Cycle_1_asus <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",experiment_date2_asus,"slopes/Cycle_1.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 

Cycle_last_asus <-read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",experiment_date2_asus,"slopes/Cycle_21.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 

Chamber volumes

chamber1_dell = 0.04650
chamber2_dell = 0.04593
chamber3_dell = 0.04977
chamber4_dell = 0.04860 

chamber1_asus = 0.04565
chamber2_asus = 0.04573
chamber3_asus = 0.04551
chamber4_asus = 0.04791

Date_tested="2023-05-16"
Clutch = "104" 
Male = "CPRE447" 
Female = "CPRE452"
Population = "Pretty patches"
Tank =171 
salinity =36 
Date_analysed = Sys.Date() 

Replicates

1

Enter specimen data

Replicate = 1 
mass = 0.0005572 
chamber = "ch4" 
Swim = "good/good"
chamber_vol = chamber4_dell
system1 = "Dell"
Notes="check max and rest"

##--- time of trail ---## 
experiment_mmr_date <- "16 May 2023 11 20AM/Oxygen"
experiment_mmr_date2 <- "16 May 2023 11 20AM/All"

firesting_mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",experiment_mmr_date,"data raw/Firesting.txt"), 
    delim = "\t", escape_double = FALSE, 
    col_types = cols(`Time (HH:MM:SS)` = col_time(format = "%H:%M:%S"), 
        `Time (s)` = col_number(), Ch1...5 = col_number(), 
        Ch2...6 = col_number(), Ch3...7 = col_number(), 
        Ch4...8 = col_number()), trim_ws = TRUE, 
    skip = 19) 
## New names:
## • `Ch1` -> `Ch1...5`
## • `Ch2` -> `Ch2...6`
## • `Ch3` -> `Ch3...7`
## • `Ch4` -> `Ch4...8`
## • `Ch 1` -> `Ch 1...9`
## • `Ch 2` -> `Ch 2...10`
## • `Ch 3` -> `Ch 3...11`
## • `Ch 4` -> `Ch 4...12`
## • `('C)` -> `('C)...15`
## • `('C)` -> `('C)...16`
## • `Ch 1` -> `Ch 1...18`
## • `Ch 2` -> `Ch 2...19`
## • `Ch 3` -> `Ch 3...20`
## • `Ch 4` -> `Ch 4...21`
## • `Ch1` -> `Ch1...22`
## • `Ch2` -> `Ch2...23`
## • `Ch3` -> `Ch3...24`
## • `Ch4` -> `Ch4...25`
## • `Ch1` -> `Ch1...26`
## • `Ch2` -> `Ch2...27`
## • `Ch3` -> `Ch3...28`
## • `Ch4` -> `Ch4...29`
## • `` -> `...31`
## Warning: One or more parsing issues, call `problems()` on your data frame for details,
## e.g.:
##   dat <- vroom(...)
##   problems(dat)
Cycle_1.mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",experiment_mmr_date2,"slopes/Cycle_1.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 
## New names:
## • `` -> `...8`

Background rates

Pre-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",preexperiment_date,"slopes")) 

pre_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

pre_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

pre_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))


bg_pre1 <- pre_cycle1 %>% calc_rate.bg()
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre2 <- pre_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre3 <- pre_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre <- mean(bg_pre1$rate.bg.mean,bg_pre2$rate.bg.mean,bg_pre3$rate.bg.mean) 
bg_pre
## [1] 1.152525e-05

post-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",postexperiment_date,"slopes")) 
 

post_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

post_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

post_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

bg_post1 <- post_cycle1 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post2 <- post_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post3 <- post_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post <- mean(bg_post1$rate.bg.mean,bg_post2$rate.bg.mean,bg_post3$rate.bg.mean)
bg_post 
## [1] -0.0009709307

Resting metabolic rate

Data manipulation

firesting2 <- firesting |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch4
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 2.18
## -----------------------------------------

#### subset data

Tstart.row=which(firesting2$TIME ==Cycle_1$Time[1], firesting$TIME) 
Tstart.dTIME=as.numeric(firesting2[Tstart.row, "dTIME"]) 

Tend.row=which(firesting2$TIME ==tail(Cycle_last$Time, n=1), firesting$TIME) 
Tend.dTIME=as.numeric(firesting2[Tend.row, "dTIME"])  

apoly_insp <- firesting2 |> 
  subset_data(from=Tstart.dTIME, 
              to=Tend.dTIME, 
              by="time") 

apoly_insp <- inspect(apoly_insp, time=1, oxygen=2)
## 
## # print.inspect # -----------------------
##                 dTIME  ch4
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8 11 13 14 15 16 17 18 23 24 25 26 27
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 2.18
## -----------------------------------------

Extract rates

apoly_cr.int <- calc_rate.int(apoly_insp, 
                              starts=(195+45+300), 
                              wait=45, 
                              measure=255, 
                              by="time", 
                              plot=TRUE) 
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from all replicates ...
## plot.calc_rate.int: Plotting first 20 selected reps only. To plot others modify 'pos' input.

## -----------------------------------------
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from selected replicates... 
## To plot others modify 'pos' input.

## -----------------------------------------

adjust rates for background

apoly_cr.int_adj <- adjust_rate(apoly_cr.int, 
                                by = bg_pre, 
                                by2 = bg_post, 
                                time_by = Tstart.row, 
                                time_by2 = Tend.row,
                                method = "linear")
## Warning: adjust_rate: One or more of the timestamps for the rate(s) in 'x' do not lie between the timestamps for the 'by' and 'by2' background rates. 
## Ensure this is correct. The adjustment value has been calculated regardless by extrapolating outside the background rates time window.
## Warning: adjust_rate: background rates in 'by' and 'by2' differ in sign (i.e. one is +ve, one is -ve). 
## Ensure this is correct. The 'linear' adjustment has been performed regardless.
## adjust_rate: Rate adjustments applied using "linear" method.
apoly_cr.int_adj$summary

Converting units

apoly_cr.int_adj2 <- apoly_cr.int_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253) 
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.
apoly_cr.int_adj2$summary

Plot curve

ggplot(as.data.frame(apoly_cr.int_adj2$summary), aes(x=row, y=rate.output*-1)) + 
  geom_point() + 
  stat_smooth(method = "lm", formula = y~poly(x, 2), color="red") +
  theme_classic()

Rate filtering

apoly_rmr <- apoly_cr.int_adj2 |> 
  select_rate(method ="rsq", n=c(0.95,1)) |> 
  select_rate(method="lowest", n=6) |> 
  plot(type="full") |> 
  summary(export = TRUE)
## select_rate: Selecting rates with rsq values between 0.95 and 1...
## ----- Selection complete. 2 rate(s) removed, 19 rate(s) remaining -----
## select_rate: Selecting lowest 6 *absolute* rate values...
## ----- Selection complete. 13 rate(s) removed, 6 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1   rsq density  row endrow     time
## 1:   7    1     279.9858 -0.01928596 0.997      NA 2977   3208  9445.71
## 2:  11    1     316.1838 -0.01875393 0.957      NA 4945   5179 11604.54
## 3:  13    1     313.2811 -0.01693879 0.985      NA 5934   6167 12684.27
## 4:  18    1     405.2419 -0.01994548 0.995      NA 8407   8641 15385.13
## 5:  19    1     405.9912 -0.01930090 0.967      NA 8900   9133 15924.38
## 6:  21    1     389.7418 -0.01710686 0.950      NA 9889  10123 17004.84
##     endtime    oxy endoxy        rate    adjustment rate.adjusted  rate.input
## 1:  9700.49 97.762 92.953 -0.01928596 -0.0003893086   -0.01889665 -0.01889665
## 2: 11859.83 98.053 93.312 -0.01875393 -0.0005988518   -0.01815508 -0.01815508
## 3: 12939.19 98.027 93.801 -0.01693879 -0.0007036237   -0.01623517 -0.01623517
## 4: 15640.79 98.095 93.251 -0.01994548 -0.0009657831   -0.01897970 -0.01897970
## 5: 16179.31 98.223 93.292 -0.01930090 -0.0010180829   -0.01828281 -0.01828281
## 6: 17260.43 98.232 93.990 -0.01710686 -0.0011229756   -0.01598388 -0.01598388
##    oxy.unit time.unit volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.0486 0.0005572   NA 36 30 1.013253 -0.2044843
## 2:     %Air       sec 0.0486 0.0005572   NA 36 30 1.013253 -0.1964596
## 3:     %Air       sec 0.0486 0.0005572   NA 36 30 1.013253 -0.1756839
## 4:     %Air       sec 0.0486 0.0005572   NA 36 30 1.013253 -0.2053830
## 5:     %Air       sec 0.0486 0.0005572   NA 36 30 1.013253 -0.1978418
## 6:     %Air       sec 0.0486 0.0005572   NA 36 30 1.013253 -0.1729647
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -366.9855          NA  mgO2/hr/kg   -366.9855
## 2:   -352.5836          NA  mgO2/hr/kg   -352.5836
## 3:   -315.2977          NA  mgO2/hr/kg   -315.2977
## 4:   -368.5983          NA  mgO2/hr/kg   -368.5983
## 5:   -355.0643          NA  mgO2/hr/kg   -355.0643
## 6:   -310.4176          NA  mgO2/hr/kg   -310.4176
## -----------------------------------------
## remove lowest slope 
apoly_rmr <- apoly_rmr |> 
  filter(rate.output != max(rate.output))

Results

results <- data.frame(Clutch = Clutch, 
                      Replicate =Replicate, 
                      Male=Male, 
                      Female=Female,
                      Population = Population, 
                      Tank = Tank,
                      Mass = mass, 
                      Chamber = chamber, 
                      System = system1,
                      Volume = chamber_vol, 
                      Date_tested = Date_tested, 
                      Date_analysed =Date_analysed,
                      Swim = Swim,
                      Salinity = salinity, 
                      Temperature = as.numeric(unique(firesting2$temperature)), 
                      Resting_kg = mean(apoly_rmr$rate.output*-1), 
                      Resting =  mean(apoly_rmr$rate.output*-1)*mass, 
                      rsqrest =mean(apoly_rmr$rsq))
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest
104 1 CPRE447 CPRE452 Pretty patches 171 0.0005572 ch4 Dell 0.0486 2023-05-16 2024-06-14 good/good 36 30 351.7059 0.1959705 0.9802

Maximum oxygen consumption

Data manipulation

firesting2_mmr <- firesting_mmr |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_mmr, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch4
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 1.70
## -----------------------------------------

Subset data

cycle1.start <-  Cycle_1.mmr[1,1]
cycle1.end <-  tail(Cycle_1.mmr, n=1)[1,1] 

cycle1.start.row <- which(firesting2_mmr$TIME == cycle1.start); cycle1.start
## Warning in which(firesting2_mmr$TIME == cycle1.start): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1.end.row <- which(firesting2_mmr$TIME == cycle1.end); cycle1.end 
## Warning in which(firesting2_mmr$TIME == cycle1.end): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1_data <- firesting2_mmr |> 
  subset_data(from = cycle1.start.row, 
              to = cycle1.end.row, 
              by = "row") 
## subset_data: Multi-column dataset detected in input! 
## subset_data is generally intended to subset data already passed through inspect(), or 2-column data frames where time and oxygen are in columns 1 and 2 respectively. 
## Subsetting will proceed anyway based on this assumption, but please ensure you understand what you are doing.
inspect(cycle1_data)
## inspect: Applying column default of 'time = 1'
## inspect: Applying column default of 'oxygen = 2'
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch4
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20 21
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.08 1.36
## -----------------------------------------

Calculating MMR

mmr <- auto_rate(cycle1_data, method = "highest", plot=TRUE, width=60, by="time") |> 
  summary()
## Warning: auto_rate: Multi-column dataset detected in input. Selecting first two columns by default.
##   If these are not the intended data, inspect() or subset the data frame columns appropriately before running auto_rate()

## 
## # summary.auto_rate # -------------------
## 
## === Summary of Results by Highest Rate ===
##      rep rank intercept_b0    slope_b1       rsq density row endrow    time
##   1:  NA    1     356.7508 -0.04576016 0.9899053      NA  54    108 5681.55
##   2:  NA    2     356.4567 -0.04570971 0.9896493      NA  53    107 5680.45
##   3:  NA    3     356.4258 -0.04570219 0.9895851      NA  55    109 5682.85
##   4:  NA    4     355.4367 -0.04552805 0.9886472      NA  56    110 5683.95
##   5:  NA    5     355.2751 -0.04550387 0.9885110      NA  52    106 5679.37
##  ---                                                                       
## 208:  NA  208     226.8004 -0.02311553 0.9798533      NA 106    160 5740.44
## 209:  NA  209     226.1128 -0.02299463 0.9843913      NA 102    156 5735.85
## 210:  NA  210     225.3528 -0.02286372 0.9837741      NA 105    159 5739.12
## 211:  NA  211     225.1963 -0.02283576 0.9844145      NA 103    157 5736.94
## 212:  NA  212     224.8351 -0.02277342 0.9846413      NA 104    158 5738.03
##      endtime    oxy endoxy        rate
##   1: 5741.55 96.586 94.157 -0.04576016
##   2: 5740.45 96.624 94.170 -0.04570971
##   3: 5742.85 96.594 94.141 -0.04570219
##   4: 5743.95 96.568 94.124 -0.04552805
##   5: 5739.37 96.653 94.205 -0.04550387
##  ---                                  
## 208: 5800.44 94.205 92.514 -0.02311553
## 209: 5795.85 94.299 92.884 -0.02299463
## 210: 5799.12 94.218 92.653 -0.02286372
## 211: 5796.94 94.268 92.848 -0.02283576
## 212: 5798.03 94.240 92.770 -0.02277342
## 
## Regressions : 212 | Results : 212 | Method : highest | Roll width : 60 | Roll type : time 
## -----------------------------------------

Adjusting

mmr_adj <- adjust_rate(mmr, by=bg_pre, method = "mean");mmr_adj
## adjust_rate: Rate adjustments applied using "mean" method.
## 
## # print.adjust_rate # -------------------
## NOTE: Consider the sign of the adjustment value when adjusting the rate.
## 
## Adjustment was applied using the 'mean' method.
## 
## Rank 1 of 212 adjusted rate(s):
## Rate          : -0.04576016
## Adjustment    : 1.152525e-05
## Adjusted Rate : -0.04577169 
## 
## To see other results use 'pos' input.
## To see full results use summary().
## -----------------------------------------

Converting units

mmr_adj2 <- mmr_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253)
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.

selecting rates

mmr_final <- mmr_adj2 |> 
  select_rate(method = "rsq", n=c(0.93,1)) |> 
  select_rate(method = "highest", n=1) |> 
  plot(type="full") |> 
  summary(export=TRUE)
## select_rate: Selecting rates with rsq values between 0.93 and 1...
## ----- Selection complete. 14 rate(s) removed, 198 rate(s) remaining -----
## select_rate: Selecting highest 1 *absolute* rate values...
## ----- Selection complete. 197 rate(s) removed, 1 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1       rsq density row endrow    time
## 1:  NA    1     356.7508 -0.04576016 0.9899053      NA  54    108 5681.55
##    endtime    oxy endoxy        rate   adjustment rate.adjusted  rate.input
## 1: 5741.55 96.586 94.157 -0.04576016 1.152525e-05   -0.04577169 -0.04577169
##    oxy.unit time.unit volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.0486 0.0005572   NA 36 30 1.013253 -0.4953042
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -888.9164          NA  mgO2/hr/kg   -888.9164
## -----------------------------------------

Results

results <-  results |> 
  mutate(Max_kg = mmr_final$rate.output*-1, 
         Max = (mmr_final$rate.output*-1)*mass, 
         rsqmax =mmr_final$rsq,
         AAS_kg = Max_kg - Resting_kg, 
         AAS = Max - Resting, 
         Notes=Notes, 
                      True_resting="") 
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest Max_kg Max rsqmax AAS_kg AAS Notes True_resting
104 1 CPRE447 CPRE452 Pretty patches 171 0.0005572 ch4 Dell 0.0486 2023-05-16 2024-06-14 good/good 36 30 351.7059 0.1959705 0.9802 888.9164 0.4953042 0.9899053 537.2105 0.2993337 check max and rest

Exporting data

resp_results_juveniles <- read_csv("resp_results_juveniles.csv") 
## Rows: 93 Columns: 25
## ── Column specification ────────────────────────────────────────────────────────
## Delimiter: ","
## chr  (9): Male, Female, Population, Chamber, System, Date_tested, Swim, Note...
## dbl (16): Clutch, Replicate, Tank, Mass, Volume, Date_analysed, Salinity, Te...
## 
## ℹ Use `spec()` to retrieve the full column specification for this data.
## ℹ Specify the column types or set `show_col_types = FALSE` to quiet this message.
resp_results_juveniles <- rbind(resp_results_juveniles, results) 
resp_results_juveniles 
write.csv(resp_results_juveniles, file="./resp_results_juveniles.csv", row.names = FALSE)

2

Enter specimen data

Replicate = 2 
mass = 0.0006846 
chamber = "ch3" 
Swim = "good/good"
chamber_vol = chamber3_dell
system1 = "Dell"
Notes="max and rest seems low"

##--- time of trail ---## 
experiment_mmr_date <- "16 May 2023 11 29AM/Oxygen"
experiment_mmr_date2 <- "16 May 2023 11 29AM/All"

firesting_mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",experiment_mmr_date,"data raw/Firesting.txt"), 
    delim = "\t", escape_double = FALSE, 
    col_types = cols(`Time (HH:MM:SS)` = col_time(format = "%H:%M:%S"), 
        `Time (s)` = col_number(), Ch1...5 = col_number(), 
        Ch2...6 = col_number(), Ch3...7 = col_number(), 
        Ch4...8 = col_number()), trim_ws = TRUE, 
    skip = 19) 
## New names:
## • `Ch1` -> `Ch1...5`
## • `Ch2` -> `Ch2...6`
## • `Ch3` -> `Ch3...7`
## • `Ch4` -> `Ch4...8`
## • `Ch 1` -> `Ch 1...9`
## • `Ch 2` -> `Ch 2...10`
## • `Ch 3` -> `Ch 3...11`
## • `Ch 4` -> `Ch 4...12`
## • `('C)` -> `('C)...15`
## • `('C)` -> `('C)...16`
## • `Ch 1` -> `Ch 1...18`
## • `Ch 2` -> `Ch 2...19`
## • `Ch 3` -> `Ch 3...20`
## • `Ch 4` -> `Ch 4...21`
## • `Ch1` -> `Ch1...22`
## • `Ch2` -> `Ch2...23`
## • `Ch3` -> `Ch3...24`
## • `Ch4` -> `Ch4...25`
## • `Ch1` -> `Ch1...26`
## • `Ch2` -> `Ch2...27`
## • `Ch3` -> `Ch3...28`
## • `Ch4` -> `Ch4...29`
## • `` -> `...31`
## Warning: One or more parsing issues, call `problems()` on your data frame for details,
## e.g.:
##   dat <- vroom(...)
##   problems(dat)
Cycle_1.mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",experiment_mmr_date2,"slopes/Cycle_1.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 
## New names:
## • `` -> `...8`

Background rates

Pre-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",preexperiment_date,"slopes")) 

pre_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

pre_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

pre_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))


bg_pre1 <- pre_cycle1 %>% calc_rate.bg()
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre2 <- pre_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre3 <- pre_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre <- mean(bg_pre1$rate.bg.mean,bg_pre2$rate.bg.mean,bg_pre3$rate.bg.mean) 
bg_pre
## [1] -3.143701e-05

post-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",postexperiment_date,"slopes")) 
 

post_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

post_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

post_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

bg_post1 <- post_cycle1 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post2 <- post_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post3 <- post_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post <- mean(bg_post1$rate.bg.mean,bg_post2$rate.bg.mean,bg_post3$rate.bg.mean)
bg_post 
## [1] -0.001637522

Resting metabolic rate

Data manipulation

firesting2 <- firesting |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch3
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 2.18
## -----------------------------------------

#### subset data

Tstart.row=which(firesting2$TIME ==Cycle_1$Time[1], firesting$TIME) 
Tstart.dTIME=as.numeric(firesting2[Tstart.row, "dTIME"]) 

Tend.row=which(firesting2$TIME ==tail(Cycle_last$Time, n=1), firesting$TIME) 
Tend.dTIME=as.numeric(firesting2[Tend.row, "dTIME"])  

apoly_insp <- firesting2 |> 
  subset_data(from=Tstart.dTIME, 
              to=Tend.dTIME, 
              by="time") 

apoly_insp <- inspect(apoly_insp, time=1, oxygen=2)
## 
## # print.inspect # -----------------------
##                 dTIME  ch3
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8 11 13 14 15 16 17 18 23 24 25 26 27
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 2.18
## -----------------------------------------

Extract rates

apoly_cr.int <- calc_rate.int(apoly_insp, 
                              starts=(195+45+300), 
                              wait=45, 
                              measure=255, 
                              by="time", 
                              plot=TRUE) 
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from all replicates ...
## plot.calc_rate.int: Plotting first 20 selected reps only. To plot others modify 'pos' input.

## -----------------------------------------
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from selected replicates... 
## To plot others modify 'pos' input.

## -----------------------------------------

adjust rates for background

apoly_cr.int_adj <- adjust_rate(apoly_cr.int, 
                                by = bg_pre, 
                                by2 = bg_post, 
                                time_by = Tstart.row, 
                                time_by2 = Tend.row,
                                method = "linear")
## Warning: adjust_rate: One or more of the timestamps for the rate(s) in 'x' do not lie between the timestamps for the 'by' and 'by2' background rates. 
## Ensure this is correct. The adjustment value has been calculated regardless by extrapolating outside the background rates time window.
## adjust_rate: Rate adjustments applied using "linear" method.
apoly_cr.int_adj$summary

Converting units

apoly_cr.int_adj2 <- apoly_cr.int_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253) 
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.
apoly_cr.int_adj2$summary

Plot curve

ggplot(as.data.frame(apoly_cr.int_adj2$summary), aes(x=row, y=rate.output*-1)) + 
  geom_point() + 
  stat_smooth(method = "lm", formula = y~poly(x, 2), color="red") +
  theme_classic()

Rate filtering

apoly_rmr <- apoly_cr.int_adj2 |> 
  select_rate(method ="rsq", n=c(0.95,1)) |> 
  select_rate(method="lowest", n=6) |> 
  plot(type="full") |> 
  summary(export = TRUE)
## select_rate: Selecting rates with rsq values between 0.95 and 1...
## ----- Selection complete. 1 rate(s) removed, 20 rate(s) remaining -----
## select_rate: Selecting lowest 6 *absolute* rate values...
## ----- Selection complete. 14 rate(s) removed, 6 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1   rsq density  row endrow     time
## 1:   6    1     325.4381 -0.02563849 0.995      NA 2488   2719  8904.94
## 2:  11    1     468.5105 -0.03200881 0.999      NA 4945   5179 11604.54
## 3:  13    1     498.8483 -0.03165654 0.999      NA 5934   6167 12684.27
## 4:  15    1     549.3055 -0.03281504 0.998      NA 6923   7157 13764.47
## 5:  16    1     563.9085 -0.03258843 0.997      NA 7417   7650 14304.28
## 6:  20    1     583.7876 -0.02949044 0.997      NA 9394   9628 16464.23
##     endtime    oxy endoxy        rate    adjustment rate.adjusted  rate.input
## 1:  9159.37 96.833 90.464 -0.02563849 -0.0006008817   -0.02503761 -0.02503761
## 2: 11859.83 97.034 88.747 -0.03200881 -0.0010292604   -0.03097955 -0.03097955
## 3: 12939.19 97.417 89.157 -0.03165654 -0.0012005378   -0.03045601 -0.03045601
## 4: 14020.18 97.616 88.957 -0.03281504 -0.0013719818   -0.03144306 -0.03144306
## 5: 14558.94 97.674 88.945 -0.03258843 -0.0014575431   -0.03113089 -0.03113089
## 6: 16719.91 98.272 90.851 -0.02949044 -0.0018003153   -0.02769012 -0.02769012
##    oxy.unit time.unit  volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.04977 0.0006846   NA 36 30 1.013253 -0.2774593
## 2:     %Air       sec 0.04977 0.0006846   NA 36 30 1.013253 -0.3433061
## 3:     %Air       sec 0.04977 0.0006846   NA 36 30 1.013253 -0.3375044
## 4:     %Air       sec 0.04977 0.0006846   NA 36 30 1.013253 -0.3484427
## 5:     %Air       sec 0.04977 0.0006846   NA 36 30 1.013253 -0.3449832
## 6:     %Air       sec 0.04977 0.0006846   NA 36 30 1.013253 -0.3068537
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -405.2868          NA  mgO2/hr/kg   -405.2868
## 2:   -501.4697          NA  mgO2/hr/kg   -501.4697
## 3:   -492.9950          NA  mgO2/hr/kg   -492.9950
## 4:   -508.9726          NA  mgO2/hr/kg   -508.9726
## 5:   -503.9194          NA  mgO2/hr/kg   -503.9194
## 6:   -448.2233          NA  mgO2/hr/kg   -448.2233
## -----------------------------------------
## remove lowest slope 
apoly_rmr <- apoly_rmr |> 
  filter(rate.output != max(rate.output))

Results

results <- data.frame(Clutch = Clutch, 
                      Replicate =Replicate, 
                      Male=Male, 
                      Female=Female,
                      Population = Population, 
                      Tank = Tank,
                      Mass = mass, 
                      Chamber = chamber, 
                      System = system1,
                      Volume = chamber_vol, 
                      Date_tested = Date_tested, 
                      Date_analysed =Date_analysed,
                      Swim = Swim,
                      Salinity = salinity, 
                      Temperature = as.numeric(unique(firesting2$temperature)), 
                      Resting_kg = mean(apoly_rmr$rate.output*-1), 
                      Resting =  mean(apoly_rmr$rate.output*-1)*mass, 
                      rsqrest =mean(apoly_rmr$rsq))
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest
104 2 CPRE447 CPRE452 Pretty patches 171 0.0006846 ch3 Dell 0.04977 2023-05-16 2024-06-14 good/good 36 30 491.116 0.336218 0.998

Maximum oxygen consumption

Data manipulation

firesting2_mmr <- firesting_mmr |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_mmr, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch3
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 2.18
## -----------------------------------------

Subset data

cycle1.start <-  Cycle_1.mmr[1,1]
cycle1.end <-  tail(Cycle_1.mmr, n=1)[1,1] 

cycle1.start.row <- which(firesting2_mmr$TIME == cycle1.start); cycle1.start
## Warning in which(firesting2_mmr$TIME == cycle1.start): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1.end.row <- which(firesting2_mmr$TIME == cycle1.end); cycle1.end 
## Warning in which(firesting2_mmr$TIME == cycle1.end): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1_data <- firesting2_mmr |> 
  subset_data(from = cycle1.start.row, 
              to = cycle1.end.row,  
              by = "row") 
## subset_data: Multi-column dataset detected in input! 
## subset_data is generally intended to subset data already passed through inspect(), or 2-column data frames where time and oxygen are in columns 1 and 2 respectively. 
## Subsetting will proceed anyway based on this assumption, but please ensure you understand what you are doing.
inspect(cycle1_data)
## inspect: Applying column default of 'time = 1'
## inspect: Applying column default of 'oxygen = 2'
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch3
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8 11 13 14 15 16 17 18 23 24 25 26 27
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.08 1.33
## -----------------------------------------

Calculating MMR

mmr <- auto_rate(cycle1_data, method = "highest", plot=TRUE, width=60, by="time") |> 
  summary()
## Warning: auto_rate: Multi-column dataset detected in input. Selecting first two columns by default.
##   If these are not the intended data, inspect() or subset the data frame columns appropriately before running auto_rate()

## 
## # summary.auto_rate # -------------------
## 
## === Summary of Results by Highest Rate ===
##      rep rank intercept_b0    slope_b1       rsq density row endrow    time
##   1:  NA    1     426.0377 -0.05261088 0.9989692      NA 158    213 6336.69
##   2:  NA    2     425.9446 -0.05259601 0.9989560      NA 159    214 6337.78
##   3:  NA    3     425.6600 -0.05255185 0.9988910      NA 157    212 6335.56
##   4:  NA    4     425.5586 -0.05253605 0.9988791      NA 156    211 6334.47
##   5:  NA    5     425.3907 -0.05250857 0.9987891      NA 160    215 6338.86
##  ---                                                                       
## 211:  NA  211     296.9231 -0.03203001 0.9639093      NA   5     60 6165.47
## 212:  NA  212     293.5017 -0.03147749 0.9641630      NA   4     59 6164.39
## 213:  NA  213     290.2563 -0.03095328 0.9645904      NA   3     58 6163.30
## 214:  NA  214     286.3943 -0.03032940 0.9652189      NA   2     57 6162.22
## 215:  NA  215     282.6021 -0.02971670 0.9657523      NA   1     56 6161.11
##      endtime    oxy endoxy        rate
##   1: 6396.69 92.723 89.537 -0.05261088
##   2: 6397.78 92.628 89.493 -0.05259601
##   3: 6395.56 92.786 89.580 -0.05255185
##   4: 6394.47 92.869 89.621 -0.05253605
##   5: 6398.86 92.602 89.497 -0.05250857
##  ---                                  
## 211: 6225.47 99.363 97.337 -0.03203001
## 212: 6224.39 99.359 97.392 -0.03147749
## 213: 6223.30 99.375 97.417 -0.03095328
## 214: 6222.22 99.404 97.477 -0.03032940
## 215: 6221.11 99.506 97.516 -0.02971670
## 
## Regressions : 215 | Results : 215 | Method : highest | Roll width : 60 | Roll type : time 
## -----------------------------------------

Adjusting

mmr_adj <- adjust_rate(mmr, by=bg_pre, method = "mean");mmr_adj
## adjust_rate: Rate adjustments applied using "mean" method.
## 
## # print.adjust_rate # -------------------
## NOTE: Consider the sign of the adjustment value when adjusting the rate.
## 
## Adjustment was applied using the 'mean' method.
## 
## Rank 1 of 215 adjusted rate(s):
## Rate          : -0.05261088
## Adjustment    : -3.143701e-05
## Adjusted Rate : -0.05257945 
## 
## To see other results use 'pos' input.
## To see full results use summary().
## -----------------------------------------

Converting units

mmr_adj2 <- mmr_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253)
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.

selecting rates

mmr_final <- mmr_adj2 |> 
  select_rate(method = "rsq", n=c(0.93,1)) |> 
  select_rate(method = "highest", n=1) |> 
  plot(type="full") |> 
  summary(export=TRUE)
## select_rate: Selecting rates with rsq values between 0.93 and 1...
## ----- Selection complete. 0 rate(s) removed, 215 rate(s) remaining -----
## select_rate: Selecting highest 1 *absolute* rate values...
## ----- Selection complete. 214 rate(s) removed, 1 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1       rsq density row endrow    time
## 1:  NA    1     426.0377 -0.05261088 0.9989692      NA 158    213 6336.69
##    endtime    oxy endoxy        rate    adjustment rate.adjusted  rate.input
## 1: 6396.69 92.723 89.537 -0.05261088 -3.143701e-05   -0.05257945 -0.05257945
##    oxy.unit time.unit  volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.04977 0.0006846   NA 36 30 1.013253 -0.5826698
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -851.1098          NA  mgO2/hr/kg   -851.1098
## -----------------------------------------

Results

results <-  results |> 
  mutate(Max_kg = mmr_final$rate.output*-1, 
         Max = (mmr_final$rate.output*-1)*mass, 
         rsqmax =mmr_final$rsq,
         AAS_kg = Max_kg - Resting_kg, 
         AAS = Max - Resting, 
                      Notes=Notes, 
                      True_resting="") 
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest Max_kg Max rsqmax AAS_kg AAS Notes True_resting
104 2 CPRE447 CPRE452 Pretty patches 171 0.0006846 ch3 Dell 0.04977 2023-05-16 2024-06-14 good/good 36 30 491.116 0.336218 0.998 851.1098 0.5826698 0.9989692 359.9938 0.2464517 max and rest seems low

Exporting data

resp_results_juveniles <- read_csv("resp_results_juveniles.csv") 
## Rows: 94 Columns: 25
## ── Column specification ────────────────────────────────────────────────────────
## Delimiter: ","
## chr  (9): Male, Female, Population, Chamber, System, Date_tested, Swim, Note...
## dbl (16): Clutch, Replicate, Tank, Mass, Volume, Date_analysed, Salinity, Te...
## 
## ℹ Use `spec()` to retrieve the full column specification for this data.
## ℹ Specify the column types or set `show_col_types = FALSE` to quiet this message.
resp_results_juveniles <- rbind(resp_results_juveniles, results) 
resp_results_juveniles 
write.csv(resp_results_juveniles, file="./resp_results_juveniles.csv", row.names = FALSE)

3

Enter specimen data

Replicate = 3 
mass = 0.0005556 
chamber = "ch2" 
Swim = "good/good"
chamber_vol = chamber2_dell
system1 = "Dell"
Notes=""

##--- time of trail ---## 
experiment_mmr_date <- "16 May 2023 11 01AM/Oxygen"
experiment_mmr_date2 <- "16 May 2023 11 01AM/All"

firesting_mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",experiment_mmr_date,"data raw/Firesting.txt"), 
    delim = "\t", escape_double = FALSE, 
    col_types = cols(`Time (HH:MM:SS)` = col_time(format = "%H:%M:%S"), 
        `Time (s)` = col_number(), Ch1...5 = col_number(), 
        Ch2...6 = col_number(), Ch3...7 = col_number(), 
        Ch4...8 = col_number()), trim_ws = TRUE, 
    skip = 19) 
## New names:
## • `Ch1` -> `Ch1...5`
## • `Ch2` -> `Ch2...6`
## • `Ch3` -> `Ch3...7`
## • `Ch4` -> `Ch4...8`
## • `Ch 1` -> `Ch 1...9`
## • `Ch 2` -> `Ch 2...10`
## • `Ch 3` -> `Ch 3...11`
## • `Ch 4` -> `Ch 4...12`
## • `('C)` -> `('C)...15`
## • `('C)` -> `('C)...16`
## • `Ch 1` -> `Ch 1...18`
## • `Ch 2` -> `Ch 2...19`
## • `Ch 3` -> `Ch 3...20`
## • `Ch 4` -> `Ch 4...21`
## • `Ch1` -> `Ch1...22`
## • `Ch2` -> `Ch2...23`
## • `Ch3` -> `Ch3...24`
## • `Ch4` -> `Ch4...25`
## • `Ch1` -> `Ch1...26`
## • `Ch2` -> `Ch2...27`
## • `Ch3` -> `Ch3...28`
## • `Ch4` -> `Ch4...29`
## • `` -> `...31`
## Warning: One or more parsing issues, call `problems()` on your data frame for details,
## e.g.:
##   dat <- vroom(...)
##   problems(dat)
Cycle_1.mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",experiment_mmr_date2,"slopes/Cycle_1.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 
## New names:
## • `` -> `...8`

Background rates

Pre-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",preexperiment_date,"slopes")) 

pre_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

pre_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

pre_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))


bg_pre1 <- pre_cycle1 %>% calc_rate.bg()
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre2 <- pre_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre3 <- pre_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre <- mean(bg_pre1$rate.bg.mean,bg_pre2$rate.bg.mean,bg_pre3$rate.bg.mean) 
bg_pre
## [1] -0.0002394606

post-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",postexperiment_date,"slopes")) 
 

post_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

post_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

post_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

bg_post1 <- post_cycle1 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post2 <- post_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post3 <- post_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post <- mean(bg_post1$rate.bg.mean,bg_post2$rate.bg.mean,bg_post3$rate.bg.mean)
bg_post 
## [1] -0.001884128

Resting metabolic rate

Data manipulation

firesting2 <- firesting |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch2
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 2.18
## -----------------------------------------

#### subset data

Tstart.row=which(firesting2$TIME ==Cycle_1$Time[1], firesting$TIME) 
Tstart.dTIME=as.numeric(firesting2[Tstart.row, "dTIME"]) 

Tend.row=which(firesting2$TIME ==tail(Cycle_last$Time, n=1), firesting$TIME) 
Tend.dTIME=as.numeric(firesting2[Tend.row, "dTIME"])  

apoly_insp <- firesting2 |> 
  subset_data(from=Tstart.dTIME, 
              to=Tend.dTIME, 
              by="time") 

apoly_insp <- inspect(apoly_insp, time=1, oxygen=2)
## 
## # print.inspect # -----------------------
##                 dTIME  ch2
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8 11 13 14 15 16 17 18 23 24 25 26 27
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 2.18
## -----------------------------------------

Extract rates

apoly_cr.int <- calc_rate.int(apoly_insp, 
                              starts=(195+45+300), 
                              wait=45, 
                              measure=255, 
                              by="time", 
                              plot=TRUE) 
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from all replicates ...
## plot.calc_rate.int: Plotting first 20 selected reps only. To plot others modify 'pos' input.

## -----------------------------------------
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from selected replicates... 
## To plot others modify 'pos' input.

## -----------------------------------------

adjust rates for background

apoly_cr.int_adj <- adjust_rate(apoly_cr.int, 
                                by = bg_pre, 
                                by2 = bg_post, 
                                time_by = Tstart.row, 
                                time_by2 = Tend.row,
                                method = "linear")
## Warning: adjust_rate: One or more of the timestamps for the rate(s) in 'x' do not lie between the timestamps for the 'by' and 'by2' background rates. 
## Ensure this is correct. The adjustment value has been calculated regardless by extrapolating outside the background rates time window.
## adjust_rate: Rate adjustments applied using "linear" method.
apoly_cr.int_adj$summary

Converting units

apoly_cr.int_adj2 <- apoly_cr.int_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253) 
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.
apoly_cr.int_adj2$summary

Plot curve

ggplot(as.data.frame(apoly_cr.int_adj2$summary), aes(x=row, y=rate.output*-1)) + 
  geom_point() + 
  stat_smooth(method = "lm", formula = y~poly(x, 2), color="red") +
  theme_classic()

Rate filtering

apoly_rmr <- apoly_cr.int_adj2 |> 
  select_rate(method ="rsq", n=c(0.95,1)) |> 
  select_rate(method="lowest", n=6) |> 
  plot(type="full") |> 
  summary(export = TRUE)
## select_rate: Selecting rates with rsq values between 0.95 and 1...
## ----- Selection complete. 2 rate(s) removed, 19 rate(s) remaining -----
## select_rate: Selecting lowest 6 *absolute* rate values...
## ----- Selection complete. 13 rate(s) removed, 6 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1   rsq density  row endrow     time
## 1:  13    1     364.8804 -0.02106375 0.982      NA 5934   6167 12684.27
## 2:  15    1     428.9900 -0.02402428 0.992      NA 6923   7157 13764.47
## 3:  16    1     444.9578 -0.02423042 0.995      NA 7417   7650 14304.28
## 4:  18    1     430.4541 -0.02163949 0.992      NA 8407   8641 15385.13
## 5:  19    1     440.1847 -0.02148718 0.991      NA 8900   9133 15924.38
## 6:  20    1     512.3845 -0.02511520 0.983      NA 9394   9628 16464.23
##     endtime    oxy endoxy        rate   adjustment rate.adjusted  rate.input
## 1: 12939.19 97.389 91.878 -0.02106375 -0.001436646   -0.01962711 -0.01962711
## 2: 14020.18 98.244 91.916 -0.02402428 -0.001612209   -0.02241207 -0.02241207
## 3: 14558.94 98.099 92.224 -0.02423042 -0.001699826   -0.02253059 -0.02253059
## 4: 15640.79 97.680 91.813 -0.02163949 -0.001875511   -0.01976398 -0.01976398
## 5: 16179.31 97.898 92.222 -0.02148718 -0.001963062   -0.01952412 -0.01952412
## 6: 16719.91 98.592 92.919 -0.02511520 -0.002050832   -0.02306437 -0.02306437
##    oxy.unit time.unit  volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.04593 0.0005556   NA 36 30 1.013253 -0.2007204
## 2:     %Air       sec 0.04593 0.0005556   NA 36 30 1.013253 -0.2292014
## 3:     %Air       sec 0.04593 0.0005556   NA 36 30 1.013253 -0.2304135
## 4:     %Air       sec 0.04593 0.0005556   NA 36 30 1.013253 -0.2021201
## 5:     %Air       sec 0.04593 0.0005556   NA 36 30 1.013253 -0.1996672
## 6:     %Air       sec 0.04593 0.0005556   NA 36 30 1.013253 -0.2358722
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -361.2679          NA  mgO2/hr/kg   -361.2679
## 2:   -412.5295          NA  mgO2/hr/kg   -412.5295
## 3:   -414.7111          NA  mgO2/hr/kg   -414.7111
## 4:   -363.7872          NA  mgO2/hr/kg   -363.7872
## 5:   -359.3722          NA  mgO2/hr/kg   -359.3722
## 6:   -424.5361          NA  mgO2/hr/kg   -424.5361
## -----------------------------------------
## remove lowest slope 
apoly_rmr <- apoly_rmr |> 
  filter(rate.output != max(rate.output))

Results

results <- data.frame(Clutch = Clutch, 
                      Replicate =Replicate, 
                      Male=Male, 
                      Female=Female,
                      Population = Population, 
                      Tank = Tank,
                      Mass = mass, 
                      Chamber = chamber, 
                      System = system1,
                      Volume = chamber_vol, 
                      Date_tested = Date_tested, 
                      Date_analysed =Date_analysed,
                      Swim = Swim,
                      Salinity = salinity, 
                      Temperature = as.numeric(unique(firesting2$temperature)), 
                      Resting_kg = mean(apoly_rmr$rate.output*-1), 
                      Resting =  mean(apoly_rmr$rate.output*-1)*mass, 
                      rsqrest =mean(apoly_rmr$rsq))
knitr::kable(results, "simple")  
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest
104 3 CPRE447 CPRE452 Pretty patches 171 0.0005556 ch2 Dell 0.04593 2023-05-16 2024-06-14 good/good 36 30 395.3663 0.2196655 0.9888

Maximum oxygen consumption

Data manipulation

firesting2_mmr <- firesting_mmr |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_mmr, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch2
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 1.70
## -----------------------------------------

Subset data

cycle1.start <-  Cycle_1.mmr[1,1]
cycle1.end <-  tail(Cycle_1.mmr, n=1)[1,1] 

cycle1.start.row <- which(firesting2_mmr$TIME == cycle1.start); cycle1.start
## Warning in which(firesting2_mmr$TIME == cycle1.start): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1.end.row <- which(firesting2_mmr$TIME == cycle1.end); cycle1.end 
## Warning in which(firesting2_mmr$TIME == cycle1.end): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1_data <- firesting2_mmr |> 
  subset_data(from = cycle1.start.row, 
              to = cycle1.end.row, 
              by = "row") 
## subset_data: Multi-column dataset detected in input! 
## subset_data is generally intended to subset data already passed through inspect(), or 2-column data frames where time and oxygen are in columns 1 and 2 respectively. 
## Subsetting will proceed anyway based on this assumption, but please ensure you understand what you are doing.
inspect(cycle1_data)
## inspect: Applying column default of 'time = 1'
## inspect: Applying column default of 'oxygen = 2'
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch2
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  2  3  4  6  7  8  9 11 12 13 14 15 16 17 18 19 20 21 22 23
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.08 1.37
## -----------------------------------------

Calculating MMR

mmr <- auto_rate(cycle1_data, method = "highest", plot=TRUE, width=60, by="time") |> 
  summary()
## Warning: auto_rate: Multi-column dataset detected in input. Selecting first two columns by default.
##   If these are not the intended data, inspect() or subset the data frame columns appropriately before running auto_rate()

## 
## # summary.auto_rate # -------------------
## 
## === Summary of Results by Highest Rate ===
##      rep rank intercept_b0    slope_b1       rsq density row endrow    time
##   1:  NA    1     417.8866 -0.07037809 0.9919310      NA 159    211 4647.37
##   2:  NA    2     417.6787 -0.07033260 0.9918311      NA 160    212 4648.46
##   3:  NA    3     417.3893 -0.07027297 0.9917123      NA 158    210 4646.19
##   4:  NA    4     417.0738 -0.07020250 0.9916210      NA 161    213 4649.79
##   5:  NA    5     416.1914 -0.07001856 0.9909555      NA 157    209 4645.10
##  ---                                                                       
## 204:  NA  204     223.6408 -0.02857423 0.9845335      NA  96    148 4574.10
## 205:  NA  205     223.5082 -0.02854719 0.9844634      NA  99    151 4577.63
## 206:  NA  206     223.3968 -0.02852186 0.9847676      NA  97    149 4575.20
## 207:  NA  207     223.3889 -0.02851931 0.9845897      NA  95    147 4572.75
## 208:  NA  208     222.8692 -0.02840783 0.9851952      NA  98    150 4576.29
##      endtime    oxy endoxy        rate
##   1: 4707.37 90.666 86.708 -0.07037809
##   2: 4708.46 90.650 86.659 -0.07033260
##   3: 4706.19 90.641 86.820 -0.07027297
##   4: 4709.79 90.625 86.566 -0.07020250
##   5: 4705.10 90.674 86.882 -0.07001856
##  ---                                  
## 204: 4634.10 92.965 91.186 -0.02857423
## 205: 4637.63 92.891 91.005 -0.02854719
## 206: 4635.20 93.005 91.134 -0.02852186
## 207: 4632.75 93.059 91.224 -0.02851931
## 208: 4636.29 92.975 91.129 -0.02840783
## 
## Regressions : 208 | Results : 208 | Method : highest | Roll width : 60 | Roll type : time 
## -----------------------------------------

Adjusting

mmr_adj <- adjust_rate(mmr, by=bg_pre, method = "mean");mmr_adj
## adjust_rate: Rate adjustments applied using "mean" method.
## 
## # print.adjust_rate # -------------------
## NOTE: Consider the sign of the adjustment value when adjusting the rate.
## 
## Adjustment was applied using the 'mean' method.
## 
## Rank 1 of 208 adjusted rate(s):
## Rate          : -0.07037809
## Adjustment    : -0.0002394606
## Adjusted Rate : -0.07013863 
## 
## To see other results use 'pos' input.
## To see full results use summary().
## -----------------------------------------

Converting units

mmr_adj2 <- mmr_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253)
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.

selecting rates

mmr_final <- mmr_adj2 |> 
  select_rate(method = "rsq", n=c(0.93,1)) |> 
  select_rate(method = "highest", n=1) |> 
  plot(type="full") |> 
  summary(export=TRUE)
## select_rate: Selecting rates with rsq values between 0.93 and 1...
## ----- Selection complete. 20 rate(s) removed, 188 rate(s) remaining -----
## select_rate: Selecting highest 1 *absolute* rate values...
## ----- Selection complete. 187 rate(s) removed, 1 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1      rsq density row endrow    time
## 1:  NA    1     417.8866 -0.07037809 0.991931      NA 159    211 4647.37
##    endtime    oxy endoxy        rate    adjustment rate.adjusted  rate.input
## 1: 4707.37 90.666 86.708 -0.07037809 -0.0002394606   -0.07013863 -0.07013863
##    oxy.unit time.unit  volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.04593 0.0005556   NA 36 30 1.013253 -0.7172864
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -1291.012          NA  mgO2/hr/kg   -1291.012
## -----------------------------------------

Results

results <-  results |> 
  mutate(Max_kg = mmr_final$rate.output*-1, 
         Max = (mmr_final$rate.output*-1)*mass, 
         rsqmax =mmr_final$rsq,
         AAS_kg = Max_kg - Resting_kg, 
         AAS = Max - Resting, 
                      Notes=Notes, 
                      True_resting="") 
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest Max_kg Max rsqmax AAS_kg AAS Notes True_resting
104 3 CPRE447 CPRE452 Pretty patches 171 0.0005556 ch2 Dell 0.04593 2023-05-16 2024-06-14 good/good 36 30 395.3663 0.2196655 0.9888 1291.012 0.7172864 0.991931 895.6459 0.4976208

Exporting data

resp_results_juveniles <- read_csv("resp_results_juveniles.csv") 
## Rows: 95 Columns: 25
## ── Column specification ────────────────────────────────────────────────────────
## Delimiter: ","
## chr  (9): Male, Female, Population, Chamber, System, Date_tested, Swim, Note...
## dbl (16): Clutch, Replicate, Tank, Mass, Volume, Date_analysed, Salinity, Te...
## 
## ℹ Use `spec()` to retrieve the full column specification for this data.
## ℹ Specify the column types or set `show_col_types = FALSE` to quiet this message.
resp_results_juveniles <- rbind(resp_results_juveniles, results) 
resp_results_juveniles 
write.csv(resp_results_juveniles, file="./resp_results_juveniles.csv", row.names = FALSE)

4

Enter specimen data

Replicate = 4 
mass = 0.0005232 
chamber = "ch1" 
Swim = "good/good"
chamber_vol = chamber1_dell
system1 = "Dell"
Notes=""

##--- time of trail ---## 
experiment_mmr_date <- "16 May 2023 11 11AM/Oxygen"
experiment_mmr_date2 <- "16 May 2023 11 11AM/All"

firesting_mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",experiment_mmr_date,"data raw/Firesting.txt"), 
    delim = "\t", escape_double = FALSE, 
    col_types = cols(`Time (HH:MM:SS)` = col_time(format = "%H:%M:%S"), 
        `Time (s)` = col_number(), Ch1...5 = col_number(), 
        Ch2...6 = col_number(), Ch3...7 = col_number(), 
        Ch4...8 = col_number()), trim_ws = TRUE, 
    skip = 19) 
## New names:
## • `Ch1` -> `Ch1...5`
## • `Ch2` -> `Ch2...6`
## • `Ch3` -> `Ch3...7`
## • `Ch4` -> `Ch4...8`
## • `Ch 1` -> `Ch 1...9`
## • `Ch 2` -> `Ch 2...10`
## • `Ch 3` -> `Ch 3...11`
## • `Ch 4` -> `Ch 4...12`
## • `('C)` -> `('C)...15`
## • `('C)` -> `('C)...16`
## • `Ch 1` -> `Ch 1...18`
## • `Ch 2` -> `Ch 2...19`
## • `Ch 3` -> `Ch 3...20`
## • `Ch 4` -> `Ch 4...21`
## • `Ch1` -> `Ch1...22`
## • `Ch2` -> `Ch2...23`
## • `Ch3` -> `Ch3...24`
## • `Ch4` -> `Ch4...25`
## • `Ch1` -> `Ch1...26`
## • `Ch2` -> `Ch2...27`
## • `Ch3` -> `Ch3...28`
## • `Ch4` -> `Ch4...29`
## • `` -> `...31`
## Warning: One or more parsing issues, call `problems()` on your data frame for details,
## e.g.:
##   dat <- vroom(...)
##   problems(dat)
Cycle_1.mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",experiment_mmr_date2,"slopes/Cycle_1.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 
## New names:
## • `` -> `...8`

Background rates

Pre-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",preexperiment_date,"slopes")) 

pre_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

pre_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

pre_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))


bg_pre1 <- pre_cycle1 %>% calc_rate.bg()
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre2 <- pre_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre3 <- pre_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre <- mean(bg_pre1$rate.bg.mean,bg_pre2$rate.bg.mean,bg_pre3$rate.bg.mean) 
bg_pre
## [1] -9.300204e-06

post-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Dell/Experiment_",postexperiment_date,"slopes")) 
 

post_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

post_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

post_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

bg_post1 <- post_cycle1 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post2 <- post_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post3 <- post_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post <- mean(bg_post1$rate.bg.mean,bg_post2$rate.bg.mean,bg_post3$rate.bg.mean)
bg_post 
## [1] -0.002014651

Resting metabolic rate

Data manipulation

firesting2 <- firesting |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch1
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 2.18
## -----------------------------------------

#### subset data

Tstart.row=which(firesting2$TIME ==Cycle_1$Time[1], firesting$TIME) 
Tstart.dTIME=as.numeric(firesting2[Tstart.row, "dTIME"]) 

Tend.row=which(firesting2$TIME ==tail(Cycle_last$Time, n=1), firesting$TIME) 
Tend.dTIME=as.numeric(firesting2[Tend.row, "dTIME"])  

apoly_insp <- firesting2 |> 
  subset_data(from=Tstart.dTIME, 
              to=Tend.dTIME, 
              by="time") 

apoly_insp <- inspect(apoly_insp, time=1, oxygen=2)
## 
## # print.inspect # -----------------------
##                 dTIME  ch1
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8 11 13 14 15 16 17 18 23 24 25 26 27
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 2.18
## -----------------------------------------

Extract rates

apoly_cr.int <- calc_rate.int(apoly_insp, 
                              starts=(195+45+300), 
                              wait=45, 
                              measure=255, 
                              by="time", 
                              plot=TRUE)  
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from all replicates ...
## plot.calc_rate.int: Plotting first 20 selected reps only. To plot others modify 'pos' input.

## -----------------------------------------
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from selected replicates... 
## To plot others modify 'pos' input.

## -----------------------------------------

adjust rates for background

apoly_cr.int_adj <- adjust_rate(apoly_cr.int, 
                                by = bg_pre, 
                                by2 = bg_post, 
                                time_by = Tstart.row, 
                                time_by2 = Tend.row,
                                method = "linear")
## Warning: adjust_rate: One or more of the timestamps for the rate(s) in 'x' do not lie between the timestamps for the 'by' and 'by2' background rates. 
## Ensure this is correct. The adjustment value has been calculated regardless by extrapolating outside the background rates time window.
## adjust_rate: Rate adjustments applied using "linear" method.
apoly_cr.int_adj$summary

Converting units

apoly_cr.int_adj2 <- apoly_cr.int_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253) 
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.
apoly_cr.int_adj2$summary

Plot curve

ggplot(as.data.frame(apoly_cr.int_adj2$summary), aes(x=row, y=rate.output*-1)) + 
  geom_point() + 
  stat_smooth(method = "lm", formula = y~poly(x, 2), color="red") +
  theme_classic()

Rate filtering

apoly_rmr <- apoly_cr.int_adj2 |> 
  select_rate(method ="rsq", n=c(0.95,1)) |> 
  select_rate(method="lowest", n=6) |> 
  plot(type="full") |> 
  summary(export = TRUE)
## select_rate: Selecting rates with rsq values between 0.95 and 1...
## ----- Selection complete. 0 rate(s) removed, 21 rate(s) remaining -----
## select_rate: Selecting lowest 6 *absolute* rate values...
## ----- Selection complete. 15 rate(s) removed, 6 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1   rsq density  row endrow     time
## 1:   8    1     345.4415 -0.02491985 0.963      NA 3467   3700  9984.93
## 2:   9    1     386.9694 -0.02759634 0.979      NA 3960   4193 10524.66
## 3:  14    1     469.0064 -0.02816033 0.997      NA 6428   6662 13224.43
## 4:  17    1     528.6197 -0.02909775 0.996      NA 7912   8146 14844.72
## 5:  20    1     574.7065 -0.02902324 0.999      NA 9394   9628 16464.23
## 6:  21    1     573.6273 -0.02804356 0.999      NA 9889  10123 17004.84
##     endtime    oxy endoxy        rate    adjustment rate.adjusted  rate.input
## 1: 10240.04 96.200 89.982 -0.02491985 -0.0009343182   -0.02398553 -0.02398553
## 2: 10780.08 96.687 88.827 -0.02759634 -0.0010412686   -0.02655507 -0.02655507
## 3: 13479.66 96.775 89.397 -0.02816033 -0.0015760702   -0.02658426 -0.02658426
## 4: 15100.18 96.460 89.106 -0.02909775 -0.0018970700   -0.02720068 -0.02720068
## 5: 16719.91 96.810 89.495 -0.02902324 -0.0022179143   -0.02680533 -0.02680533
## 6: 17260.43 96.951 89.402 -0.02804356 -0.0023249994   -0.02571856 -0.02571856
##    oxy.unit time.unit volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.0465 0.0005232   NA 36 30 1.013253 -0.2483368
## 2:     %Air       sec 0.0465 0.0005232   NA 36 30 1.013253 -0.2749408
## 3:     %Air       sec 0.0465 0.0005232   NA 36 30 1.013253 -0.2752431
## 4:     %Air       sec 0.0465 0.0005232   NA 36 30 1.013253 -0.2816252
## 5:     %Air       sec 0.0465 0.0005232   NA 36 30 1.013253 -0.2775319
## 6:     %Air       sec 0.0465 0.0005232   NA 36 30 1.013253 -0.2662800
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -474.6499          NA  mgO2/hr/kg   -474.6499
## 2:   -525.4985          NA  mgO2/hr/kg   -525.4985
## 3:   -526.0762          NA  mgO2/hr/kg   -526.0762
## 4:   -538.2745          NA  mgO2/hr/kg   -538.2745
## 5:   -530.4509          NA  mgO2/hr/kg   -530.4509
## 6:   -508.9449          NA  mgO2/hr/kg   -508.9449
## -----------------------------------------
## remove lowest slope 
apoly_rmr <- apoly_rmr |> 
  filter(rate.output != max(rate.output))

Results

results <- data.frame(Clutch = Clutch, 
                      Replicate =Replicate, 
                      Male=Male, 
                      Female=Female,
                      Population = Population, 
                      Tank = Tank,
                      Mass = mass, 
                      Chamber = chamber, 
                      System = system1,
                      Volume = chamber_vol, 
                      Date_tested = Date_tested, 
                      Date_analysed =Date_analysed,
                      Swim = Swim,
                      Salinity = salinity, 
                      Temperature = as.numeric(unique(firesting2$temperature)), 
                      Resting_kg = mean(apoly_rmr$rate.output*-1), 
                      Resting =  mean(apoly_rmr$rate.output*-1)*mass, 
                      rsqrest =mean(apoly_rmr$rsq))
knitr::kable(results, "simple")  
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest
104 4 CPRE447 CPRE452 Pretty patches 171 0.0005232 ch1 Dell 0.0465 2023-05-16 2024-06-14 good/good 36 30 525.849 0.2751242 0.994

Maximum oxygen consumption

Data manipulation

firesting2_mmr <- firesting_mmr |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_mmr, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch1
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.07 1.70
## -----------------------------------------

Subset data

cycle1.start <-  Cycle_1.mmr[1,1]
cycle1.end <-  tail(Cycle_1.mmr, n=1)[1,1] 

cycle1.start.row <- which(firesting2_mmr$TIME == cycle1.start); cycle1.start
## Warning in which(firesting2_mmr$TIME == cycle1.start): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1.end.row <- which(firesting2_mmr$TIME == cycle1.end); cycle1.end 
## Warning in which(firesting2_mmr$TIME == cycle1.end): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1_data <- firesting2_mmr |> 
  subset_data(from = cycle1.start.row, 
              to = cycle1.end.row, 
              by = "row") 
## subset_data: Multi-column dataset detected in input! 
## subset_data is generally intended to subset data already passed through inspect(), or 2-column data frames where time and oxygen are in columns 1 and 2 respectively. 
## Subsetting will proceed anyway based on this assumption, but please ensure you understand what you are doing.
inspect(cycle1_data)
## inspect: Applying column default of 'time = 1'
## inspect: Applying column default of 'oxygen = 2'
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch1
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  6  7  8  9 10 12 13 14 15 16 17 19 20 21 22 24
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.08 1.35
## -----------------------------------------

Calculating MMR

mmr <- auto_rate(cycle1_data, method = "highest", plot=TRUE, width=60, by="time") |> 
  summary()
## Warning: auto_rate: Multi-column dataset detected in input. Selecting first two columns by default.
##   If these are not the intended data, inspect() or subset the data frame columns appropriately before running auto_rate()

## 
## # summary.auto_rate # -------------------
## 
## === Summary of Results by Highest Rate ===
##      rep rank intercept_b0    slope_b1       rsq density row endrow    time
##   1:  NA    1     450.6658 -0.06901485 0.9933309      NA  45     98 5119.71
##   2:  NA    2     450.3445 -0.06895337 0.9932102      NA  44     97 5118.39
##   3:  NA    3     450.2617 -0.06893525 0.9931498      NA  46     99 5120.81
##   4:  NA    4     449.7439 -0.06883732 0.9930565      NA  43     96 5117.29
##   5:  NA    5     449.3989 -0.06877063 0.9930011      NA  42     95 5116.19
##  ---                                                                       
## 206:  NA  206     247.9367 -0.02982536 0.9665733      NA  87    140 5167.96
## 207:  NA  207     245.5055 -0.02935671 0.9679017      NA  86    139 5166.86
## 208:  NA  208     244.6259 -0.02918392 0.9693475      NA  83    136 5163.60
## 209:  NA  209     244.1519 -0.02909506 0.9704385      NA  85    138 5165.77
## 210:  NA  210     243.7288 -0.02901251 0.9712124      NA  84    137 5164.68
##      endtime    oxy endoxy        rate
##   1: 5179.71 97.300 93.295 -0.06901485
##   2: 5178.39 97.426 93.328 -0.06895337
##   3: 5180.81 97.203 93.290 -0.06893525
##   4: 5177.29 97.501 93.380 -0.06883732
##   5: 5176.19 97.572 93.438 -0.06877063
##  ---                                  
## 206: 5227.96 93.914 91.771 -0.02982536
## 207: 5226.86 93.987 91.842 -0.02935671
## 208: 5223.60 94.339 92.098 -0.02918392
## 209: 5225.77 94.114 91.938 -0.02909506
## 210: 5224.68 94.204 92.021 -0.02901251
## 
## Regressions : 210 | Results : 210 | Method : highest | Roll width : 60 | Roll type : time 
## -----------------------------------------

Adjusting

mmr_adj <- adjust_rate(mmr, by=bg_pre, method = "mean");mmr_adj
## adjust_rate: Rate adjustments applied using "mean" method.
## 
## # print.adjust_rate # -------------------
## NOTE: Consider the sign of the adjustment value when adjusting the rate.
## 
## Adjustment was applied using the 'mean' method.
## 
## Rank 1 of 210 adjusted rate(s):
## Rate          : -0.06901485
## Adjustment    : -9.300204e-06
## Adjusted Rate : -0.06900555 
## 
## To see other results use 'pos' input.
## To see full results use summary().
## -----------------------------------------

Converting units

mmr_adj2 <- mmr_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253)
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.

selecting rates

mmr_final <- mmr_adj2 |> 
  select_rate(method = "rsq", n=c(0.93,1)) |> 
  select_rate(method = "highest", n=1) |> 
  plot(type="full") |> 
  summary(export=TRUE)
## select_rate: Selecting rates with rsq values between 0.93 and 1...
## ----- Selection complete. 16 rate(s) removed, 194 rate(s) remaining -----
## select_rate: Selecting highest 1 *absolute* rate values...
## ----- Selection complete. 193 rate(s) removed, 1 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1       rsq density row endrow    time
## 1:  NA    1     450.6658 -0.06901485 0.9933309      NA  45     98 5119.71
##    endtime  oxy endoxy        rate    adjustment rate.adjusted  rate.input
## 1: 5179.71 97.3 93.295 -0.06901485 -9.300204e-06   -0.06900555 -0.06900555
##    oxy.unit time.unit volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.0465 0.0005232   NA 36 30 1.013253 -0.7144565
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -1365.551          NA  mgO2/hr/kg   -1365.551
## -----------------------------------------

Results

results <-  results |> 
  mutate(Max_kg = mmr_final$rate.output*-1, 
         Max = (mmr_final$rate.output*-1)*mass, 
         rsqmax =mmr_final$rsq,
         AAS_kg = Max_kg - Resting_kg, 
         AAS = Max - Resting, 
                      Notes=Notes, 
                      True_resting="") 
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest Max_kg Max rsqmax AAS_kg AAS Notes True_resting
104 4 CPRE447 CPRE452 Pretty patches 171 0.0005232 ch1 Dell 0.0465 2023-05-16 2024-06-14 good/good 36 30 525.849 0.2751242 0.994 1365.551 0.7144565 0.9933309 839.7025 0.4393323

Exporting data

resp_results_juveniles <- read_csv("resp_results_juveniles.csv") 
## Rows: 96 Columns: 25
## ── Column specification ────────────────────────────────────────────────────────
## Delimiter: ","
## chr  (9): Male, Female, Population, Chamber, System, Date_tested, Swim, Note...
## dbl (16): Clutch, Replicate, Tank, Mass, Volume, Date_analysed, Salinity, Te...
## 
## ℹ Use `spec()` to retrieve the full column specification for this data.
## ℹ Specify the column types or set `show_col_types = FALSE` to quiet this message.
resp_results_juveniles <- rbind(resp_results_juveniles, results) 
resp_results_juveniles 
write.csv(resp_results_juveniles, file="./resp_results_juveniles.csv", row.names = FALSE)

5

Enter specimen data

Replicate = 5 
mass = 0.0008584 
chamber = "ch4" 
Swim = "good/good"
chamber_vol = chamber4_asus
system1 = "Asus"
Notes=""

##--- time of trail ---## 
experiment_mmr_date_asus <- "16 May 2023 12 00PM/Oxygen"
experiment_mmr_date2_asus <- "16 May 2023 12 00PM/All"

firesting_mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",experiment_mmr_date_asus,"data raw/Firesting.txt"), 
    delim = "\t", escape_double = FALSE, 
    col_types = cols(`Time (HH:MM:SS)` = col_time(format = "%H:%M:%S"), 
        `Time (s)` = col_number(), Ch1...5 = col_number(), 
        Ch2...6 = col_number(), Ch3...7 = col_number(), 
        Ch4...8 = col_number()), trim_ws = TRUE, 
    skip = 19) 
## New names:
## • `Ch1` -> `Ch1...5`
## • `Ch2` -> `Ch2...6`
## • `Ch3` -> `Ch3...7`
## • `Ch4` -> `Ch4...8`
## • `Ch 1` -> `Ch 1...9`
## • `Ch 2` -> `Ch 2...10`
## • `Ch 3` -> `Ch 3...11`
## • `Ch 4` -> `Ch 4...12`
## • `('C)` -> `('C)...15`
## • `('C)` -> `('C)...16`
## • `Ch 1` -> `Ch 1...18`
## • `Ch 2` -> `Ch 2...19`
## • `Ch 3` -> `Ch 3...20`
## • `Ch 4` -> `Ch 4...21`
## • `Ch1` -> `Ch1...22`
## • `Ch2` -> `Ch2...23`
## • `Ch3` -> `Ch3...24`
## • `Ch4` -> `Ch4...25`
## • `Ch1` -> `Ch1...26`
## • `Ch2` -> `Ch2...27`
## • `Ch3` -> `Ch3...28`
## • `Ch4` -> `Ch4...29`
## • `` -> `...31`
## Warning: One or more parsing issues, call `problems()` on your data frame for details,
## e.g.:
##   dat <- vroom(...)
##   problems(dat)
Cycle_1.mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",experiment_mmr_date2_asus,"slopes/Cycle_1.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 
## New names:
## • `` -> `...8`

Background rates

Pre-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",preexperiment_date_asus,"slopes")) 

pre_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

pre_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

pre_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))


bg_pre1 <- pre_cycle1 %>% calc_rate.bg()
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre2 <- pre_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre3 <- pre_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre <- mean(bg_pre1$rate.bg.mean,bg_pre2$rate.bg.mean,bg_pre3$rate.bg.mean) 
bg_pre
## [1] 0.0009788346

post-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",postexperiment_date_asus,"slopes")) 
 

post_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

post_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

post_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

bg_post1 <- post_cycle1 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post2 <- post_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post3 <- post_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post <- mean(bg_post1$rate.bg.mean,bg_post2$rate.bg.mean,bg_post3$rate.bg.mean)
bg_post 
## [1] -0.0006355945

Resting metabolic rate

Data manipulation

firesting2_asus <- firesting_asus |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_asus, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch4
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 5.08
## -----------------------------------------

#### subset data

Tstart.row=which(firesting2_asus$TIME ==Cycle_1_asus$Time[1], firesting_asus$TIME) 
Tstart.dTIME=as.numeric(firesting2_asus[Tstart.row, "dTIME"]) 

Tend.row=which(firesting2_asus$TIME ==tail(Cycle_last_asus$Time, n=1), firesting_asus$TIME) 
Tend.dTIME=as.numeric(firesting2_asus[Tend.row, "dTIME"])  

apoly_insp <- firesting2_asus |> 
  subset_data(from=Tstart.dTIME, 
              to=Tend.dTIME, 
              by="time") 

apoly_insp <- inspect(apoly_insp, time=1, oxygen=2)
## 
## # print.inspect # -----------------------
##                 dTIME  ch4
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  4  5  6  7  8  9 10 11 12 13 14 15 16 17 19 20 21 23
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 5.08
## -----------------------------------------

Extract rates

apoly_cr.int <- calc_rate.int(apoly_insp, 
                              starts=(195+45+300), 
                              wait=45, 
                              measure=255, 
                              by="time", 
                              plot=TRUE) 
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from all replicates ...
## plot.calc_rate.int: Plotting first 20 selected reps only. To plot others modify 'pos' input.

## -----------------------------------------

adjust rates for background

apoly_cr.int_adj <- adjust_rate(apoly_cr.int, 
                                by = bg_pre, 
                                by2 = bg_post, 
                                time_by = Tstart.row, 
                                time_by2 = Tend.row,
                                method = "linear")
## Warning: adjust_rate: One or more of the timestamps for the rate(s) in 'x' do not lie between the timestamps for the 'by' and 'by2' background rates. 
## Ensure this is correct. The adjustment value has been calculated regardless by extrapolating outside the background rates time window.
## Warning: adjust_rate: background rates in 'by' and 'by2' differ in sign (i.e. one is +ve, one is -ve). 
## Ensure this is correct. The 'linear' adjustment has been performed regardless.
## adjust_rate: Rate adjustments applied using "linear" method.
apoly_cr.int_adj$summary

Converting units

apoly_cr.int_adj2 <- apoly_cr.int_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253) 
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.
apoly_cr.int_adj2$summary

Plot curve

ggplot(as.data.frame(apoly_cr.int_adj2$summary), aes(x=row, y=rate.output*-1)) + 
  geom_point() + 
  stat_smooth(method = "lm", formula = y~poly(x, 2), color="red") +
  theme_classic()

Rate filtering

apoly_rmr <- apoly_cr.int_adj2 |> 
  select_rate(method ="rsq", n=c(0.95,1)) |> 
  select_rate(method="lowest", n=6) |> 
  plot(type="full") |> 
  summary(export = TRUE)
## select_rate: Selecting rates with rsq values between 0.95 and 1...
## ----- Selection complete. 0 rate(s) removed, 21 rate(s) remaining -----
## select_rate: Selecting lowest 6 *absolute* rate values...
## ----- Selection complete. 15 rate(s) removed, 6 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1   rsq density  row endrow     time
## 1:   9    1     537.0789 -0.03458594 0.996      NA 3211   3399 12679.69
## 2:  10    1     548.0261 -0.03403614 0.992      NA 3608   3773 13219.12
## 3:  11    1     564.8386 -0.03383623 0.997      NA 3904   4049 13759.09
## 4:  15    1     656.3794 -0.03500099 0.993      NA 5453   5640 15919.70
## 5:  16    1     666.6968 -0.03448608 0.998      NA 5850   6038 16458.92
## 6:  18    1     673.1659 -0.03278989 0.986      NA 6645   6833 17539.79
##     endtime    oxy endoxy        rate    adjustment rate.adjusted  rate.input
## 1: 12935.07 98.214 89.638 -0.03458594 -0.0003695143   -0.03421643 -0.03421643
## 2: 13473.98 98.057 89.064 -0.03403614 -0.0004779007   -0.03355824 -0.03355824
## 3: 14014.67 98.979 90.589 -0.03383623 -0.0005865204   -0.03324971 -0.03324971
## 4: 16174.14 98.783 90.225 -0.03500099 -0.0010207417   -0.03398025 -0.03398025
## 5: 16714.49 98.810 90.138 -0.03448608 -0.0011292518   -0.03335683 -0.03335683
## 6: 17795.32 97.969 89.004 -0.03278989 -0.0013465294   -0.03144336 -0.03144336
##    oxy.unit time.unit  volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.04791 0.0008584   NA 36 30 1.013253 -0.3650057
## 2:     %Air       sec 0.04791 0.0008584   NA 36 30 1.013253 -0.3579845
## 3:     %Air       sec 0.04791 0.0008584   NA 36 30 1.013253 -0.3546932
## 4:     %Air       sec 0.04791 0.0008584   NA 36 30 1.013253 -0.3624862
## 5:     %Air       sec 0.04791 0.0008584   NA 36 30 1.013253 -0.3558359
## 6:     %Air       sec 0.04791 0.0008584   NA 36 30 1.013253 -0.3354239
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -425.2164          NA  mgO2/hr/kg   -425.2164
## 2:   -417.0369          NA  mgO2/hr/kg   -417.0369
## 3:   -413.2027          NA  mgO2/hr/kg   -413.2027
## 4:   -422.2813          NA  mgO2/hr/kg   -422.2813
## 5:   -414.5339          NA  mgO2/hr/kg   -414.5339
## 6:   -390.7548          NA  mgO2/hr/kg   -390.7548
## -----------------------------------------
## remove lowest slope 
apoly_rmr <- apoly_rmr |> 
  filter(rate.output != max(rate.output))

Results

results <- data.frame(Clutch = Clutch, 
                      Replicate =Replicate, 
                      Male=Male, 
                      Female=Female,
                      Population = Population, 
                      Tank = Tank,
                      Mass = mass, 
                      Chamber = chamber, 
                      System = system1,
                      Volume = chamber_vol, 
                      Date_tested = Date_tested, 
                      Date_analysed =Date_analysed,
                      Swim = Swim,
                      Salinity = salinity, 
                      Temperature = as.numeric(unique(firesting2$temperature)), 
                      Resting_kg = mean(apoly_rmr$rate.output*-1), 
                      Resting =  mean(apoly_rmr$rate.output*-1)*mass, 
                      rsqrest =mean(apoly_rmr$rsq))
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest
104 5 CPRE447 CPRE452 Pretty patches 171 0.0008584 ch4 Asus 0.04791 2023-05-16 2024-06-14 good/good 36 30 418.4542 0.3592011 0.9952

Maximum oxygen consumption

Data manipulation

firesting2_mmr <- firesting_mmr |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_mmr, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch4
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 1.87
## -----------------------------------------

Subset data

cycle1.start <-  Cycle_1.mmr[1,1]
cycle1.end <-  tail(Cycle_1.mmr, n=1)[1,1] 

cycle1.start.row <- which(firesting2_mmr$TIME == cycle1.start); cycle1.start
## Warning in which(firesting2_mmr$TIME == cycle1.start): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1.end.row <- which(firesting2_mmr$TIME == cycle1.end); cycle1.end 
## Warning in which(firesting2_mmr$TIME == cycle1.end): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1_data <- firesting2_mmr |> 
  subset_data(from = cycle1.start.row, 
              to = cycle1.end.row, 
              by = "row") 
## subset_data: Multi-column dataset detected in input! 
## subset_data is generally intended to subset data already passed through inspect(), or 2-column data frames where time and oxygen are in columns 1 and 2 respectively. 
## Subsetting will proceed anyway based on this assumption, but please ensure you understand what you are doing.
inspect(cycle1_data)
## inspect: Applying column default of 'time = 1'
## inspect: Applying column default of 'oxygen = 2'
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch4
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 12 13 14 15 17 18 19 20 21 22
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 1.68
## -----------------------------------------

Calculating MMR

mmr <- auto_rate(cycle1_data, method = "highest", plot=TRUE, width=60, by="time") |> 
  summary()
## Warning: auto_rate: Multi-column dataset detected in input. Selecting first two columns by default.
##   If these are not the intended data, inspect() or subset the data frame columns appropriately before running auto_rate()

## 
## # summary.auto_rate # -------------------
## 
## === Summary of Results by Highest Rate ===
##      rep rank intercept_b0    slope_b1       rsq density row endrow    time
##   1:  NA    1     822.5337 -0.11952227 0.9916584      NA  54     99 6097.60
##   2:  NA    2     822.2128 -0.11946835 0.9915973      NA  55    100 6098.97
##   3:  NA    3     821.0118 -0.11927523 0.9914169      NA  53     98 6096.24
##   4:  NA    4     820.9362 -0.11925810 0.9913092      NA  56    101 6100.33
##   5:  NA    5     818.2014 -0.11881748 0.9910558      NA  52     97 6094.89
##  ---                                                                       
## 172:  NA  172     235.0882 -0.02488765 0.6091335      NA 172    217 6258.87
## 173:  NA  173     226.6806 -0.02355056 0.5785091      NA 173    218 6260.23
## 174:  NA  174     220.3181 -0.02253923 0.5562255      NA 174    219 6261.63
## 175:  NA  175     216.1174 -0.02187198 0.5420465      NA 175    220 6262.96
## 176:  NA  176     212.7148 -0.02133203 0.5312055      NA 176    221 6264.33
##      endtime    oxy endoxy        rate
##   1: 6157.60 93.628 86.693 -0.11952227
##   2: 6158.97 93.518 86.634 -0.11946835
##   3: 6156.24 93.768 86.766 -0.11927523
##   4: 6160.33 93.340 86.578 -0.11925810
##   5: 6154.89 93.815 86.861 -0.11881748
##  ---                                  
## 172: 6318.87 79.700 78.247 -0.02488765
## 173: 6320.23 79.604 78.176 -0.02355056
## 174: 6321.63 79.496 78.050 -0.02253923
## 175: 6322.96 79.416 77.921 -0.02187198
## 176: 6324.33 79.331 77.829 -0.02133203
## 
## Regressions : 176 | Results : 176 | Method : highest | Roll width : 60 | Roll type : time 
## -----------------------------------------

Adjusting

mmr_adj <- adjust_rate(mmr, by=bg_pre, method = "mean");mmr_adj
## adjust_rate: Rate adjustments applied using "mean" method.
## 
## # print.adjust_rate # -------------------
## NOTE: Consider the sign of the adjustment value when adjusting the rate.
## 
## Adjustment was applied using the 'mean' method.
## 
## Rank 1 of 176 adjusted rate(s):
## Rate          : -0.1195223
## Adjustment    : 0.0009788346
## Adjusted Rate : -0.1205011 
## 
## To see other results use 'pos' input.
## To see full results use summary().
## -----------------------------------------

Converting units

mmr_adj2 <- mmr_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253)
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.

selecting rates

mmr_final <- mmr_adj2 |> 
  select_rate(method = "rsq", n=c(0.93,1)) |> 
  select_rate(method = "highest", n=1) |> 
  plot(type="full") |> 
  summary(export=TRUE)
## select_rate: Selecting rates with rsq values between 0.93 and 1...
## ----- Selection complete. 32 rate(s) removed, 144 rate(s) remaining -----
## select_rate: Selecting highest 1 *absolute* rate values...
## ----- Selection complete. 143 rate(s) removed, 1 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0   slope_b1       rsq density row endrow   time endtime
## 1:  NA    1     822.5337 -0.1195223 0.9916584      NA  54     99 6097.6  6157.6
##       oxy endoxy       rate   adjustment rate.adjusted rate.input oxy.unit
## 1: 93.628 86.693 -0.1195223 0.0009788346    -0.1205011 -0.1205011     %Air
##    time.unit  volume      mass area  S  t        P  rate.abs rate.m.spec
## 1:       sec 0.04791 0.0008584   NA 36 30 1.013253 -1.285453   -1497.498
##    rate.a.spec output.unit rate.output
## 1:          NA  mgO2/hr/kg   -1497.498
## -----------------------------------------

Results

results <-  results |> 
  mutate(Max_kg = mmr_final$rate.output*-1, 
         Max = (mmr_final$rate.output*-1)*mass,
         rsqmax =mmr_final$rsq,
         AAS_kg = Max_kg - Resting_kg, 
         AAS = Max - Resting, 
         Notes=Notes, 
         True_resting="") 
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest Max_kg Max rsqmax AAS_kg AAS Notes True_resting
104 5 CPRE447 CPRE452 Pretty patches 171 0.0008584 ch4 Asus 0.04791 2023-05-16 2024-06-14 good/good 36 30 418.4542 0.3592011 0.9952 1497.498 1.285453 0.9916584 1079.044 0.9262514
### Expor ting data
resp_results_juveniles <- read_csv("resp_results_juveniles.csv") 
## Rows: 97 Columns: 25
## ── Column specification ────────────────────────────────────────────────────────
## Delimiter: ","
## chr  (9): Male, Female, Population, Chamber, System, Date_tested, Swim, Note...
## dbl (16): Clutch, Replicate, Tank, Mass, Volume, Date_analysed, Salinity, Te...
## 
## ℹ Use `spec()` to retrieve the full column specification for this data.
## ℹ Specify the column types or set `show_col_types = FALSE` to quiet this message.
resp_results_juveniles <- rbind(resp_results_juveniles, results) 
resp_results_juveniles 
write.csv(resp_results_juveniles, file="./resp_results_juveniles.csv", row.names = FALSE)

6

Enter specimen data

Replicate = 6 
mass = 0.0004818 
chamber = "ch3" 
Swim = "good/good"
chamber_vol = chamber3_asus
system1 = "Asus"
Notes=""

##--- time of trail ---## 
experiment_mmr_date_asus <- "16 May 2023 12 17PM/Oxygen"
experiment_mmr_date2_asus <- "16 May 2023 12 17PM/All"

firesting_mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",experiment_mmr_date_asus,"data raw/Firesting.txt"), 
    delim = "\t", escape_double = FALSE, 
    col_types = cols(`Time (HH:MM:SS)` = col_time(format = "%H:%M:%S"), 
        `Time (s)` = col_number(), Ch1...5 = col_number(), 
        Ch2...6 = col_number(), Ch3...7 = col_number(), 
        Ch4...8 = col_number()), trim_ws = TRUE, 
    skip = 19) 
## New names:
## • `Ch1` -> `Ch1...5`
## • `Ch2` -> `Ch2...6`
## • `Ch3` -> `Ch3...7`
## • `Ch4` -> `Ch4...8`
## • `Ch 1` -> `Ch 1...9`
## • `Ch 2` -> `Ch 2...10`
## • `Ch 3` -> `Ch 3...11`
## • `Ch 4` -> `Ch 4...12`
## • `('C)` -> `('C)...15`
## • `('C)` -> `('C)...16`
## • `Ch 1` -> `Ch 1...18`
## • `Ch 2` -> `Ch 2...19`
## • `Ch 3` -> `Ch 3...20`
## • `Ch 4` -> `Ch 4...21`
## • `Ch1` -> `Ch1...22`
## • `Ch2` -> `Ch2...23`
## • `Ch3` -> `Ch3...24`
## • `Ch4` -> `Ch4...25`
## • `Ch1` -> `Ch1...26`
## • `Ch2` -> `Ch2...27`
## • `Ch3` -> `Ch3...28`
## • `Ch4` -> `Ch4...29`
## • `` -> `...31`
## Warning: One or more parsing issues, call `problems()` on your data frame for details,
## e.g.:
##   dat <- vroom(...)
##   problems(dat)
Cycle_1.mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",experiment_mmr_date2_asus,"slopes/Cycle_1.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 
## New names:
## • `` -> `...8`

Background rates

Pre-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",preexperiment_date_asus,"slopes")) 

pre_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

pre_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

pre_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))


bg_pre1 <- pre_cycle1 %>% calc_rate.bg()
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre2 <- pre_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre3 <- pre_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre <- mean(bg_pre1$rate.bg.mean,bg_pre2$rate.bg.mean,bg_pre3$rate.bg.mean) 
bg_pre
## [1] -0.0003964544

post-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",postexperiment_date_asus,"slopes")) 
 

post_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

post_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

post_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

bg_post1 <- post_cycle1 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post2 <- post_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post3 <- post_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post <- mean(bg_post1$rate.bg.mean,bg_post2$rate.bg.mean,bg_post3$rate.bg.mean)
bg_post 
## [1] -0.00164869

Resting metabolic rate

Data manipulation

firesting2_asus <- firesting_asus |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_asus, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch3
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 5.08
## -----------------------------------------

#### subset data

Tstart.row=which(firesting2_asus$TIME ==Cycle_1_asus$Time[1], firesting_asus$TIME) 
Tstart.dTIME=as.numeric(firesting2_asus[Tstart.row, "dTIME"]) 

Tend.row=which(firesting2_asus$TIME ==tail(Cycle_last_asus$Time, n=1), firesting_asus$TIME) 
Tend.dTIME=as.numeric(firesting2_asus[Tend.row, "dTIME"])  

apoly_insp <- firesting2_asus |> 
  subset_data(from=Tstart.dTIME, 
              to=Tend.dTIME, 
              by="time") 

apoly_insp <- inspect(apoly_insp, time=1, oxygen=2)
## 
## # print.inspect # -----------------------
##                 dTIME  ch3
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  4  5  6  7  8  9 10 11 12 13 14 15 16 17 19 20 21 23
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 5.08
## -----------------------------------------

Extract rates

apoly_cr.int <- calc_rate.int(apoly_insp, 
                              starts=(195+45+300), 
                              wait=45, 
                              measure=245, 
                              by="time", 
                              plot=TRUE) 
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from all replicates ...
## plot.calc_rate.int: Plotting first 20 selected reps only. To plot others modify 'pos' input.

## -----------------------------------------

adjust rates for background

apoly_cr.int_adj <- adjust_rate(apoly_cr.int, 
                                by = bg_pre, 
                                by2 = bg_post, 
                                time_by = Tstart.row, 
                                time_by2 = Tend.row,
                                method = "linear")
## Warning: adjust_rate: One or more of the timestamps for the rate(s) in 'x' do not lie between the timestamps for the 'by' and 'by2' background rates. 
## Ensure this is correct. The adjustment value has been calculated regardless by extrapolating outside the background rates time window.
## adjust_rate: Rate adjustments applied using "linear" method.
apoly_cr.int_adj$summary

Converting units

apoly_cr.int_adj2 <- apoly_cr.int_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253) 
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.
apoly_cr.int_adj2$summary

Plot curve

ggplot(as.data.frame(apoly_cr.int_adj2$summary), aes(x=row, y=rate.output*-1)) + 
  geom_point() + 
  stat_smooth(method = "lm", formula = y~poly(x, 2), color="red") +
  theme_classic()

Rate filtering

apoly_rmr <- apoly_cr.int_adj2 |> 
  select_rate(method ="rsq", n=c(0.95,1)) |> 
  select_rate(method="lowest", n=6) |> 
  plot(type="full") |> 
  summary(export = TRUE)
## select_rate: Selecting rates with rsq values between 0.95 and 1...
## ----- Selection complete. 1 rate(s) removed, 20 rate(s) remaining -----
## select_rate: Selecting lowest 6 *absolute* rate values...
## ----- Selection complete. 14 rate(s) removed, 6 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1   rsq density  row endrow     time
## 1:   6    1     376.7217 -0.02524086 0.990      NA 2018   2198 11058.76
## 2:   9    1     398.3244 -0.02374681 0.991      NA 3211   3391 12679.69
## 3:  10    1     418.1756 -0.02424000 0.970      NA 3608   3770 13219.12
## 4:  18    1     543.6151 -0.02539817 0.978      NA 6645   6825 17539.79
## 5:  19    1     550.1205 -0.02499908 0.994      NA 7042   7223 18079.16
## 6:  21    1     534.5721 -0.02283163 0.989      NA 7837   8018 19159.00
##     endtime    oxy endoxy        rate   adjustment rate.adjusted  rate.input
## 1: 11303.55 97.885 91.457 -0.02524086 -0.001188735   -0.02405212 -0.02405212
## 2: 12924.12 97.214 91.766 -0.02374681 -0.001441451   -0.02230535 -0.02230535
## 3: 13463.66 97.497 91.181 -0.02424000 -0.001525570   -0.02271443 -0.02271443
## 4: 17784.46 97.633 91.527 -0.02539817 -0.002199281   -0.02319889 -0.02319889
## 5: 18324.95 97.847 92.233 -0.02499908 -0.002283470   -0.02271561 -0.02271561
## 6: 19404.90 97.267 91.508 -0.02283163 -0.002451853   -0.02037978 -0.02037978
##    oxy.unit time.unit  volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.04551 0.0004818   NA 36 30 1.013253 -0.2437244
## 2:     %Air       sec 0.04551 0.0004818   NA 36 30 1.013253 -0.2260241
## 3:     %Air       sec 0.04551 0.0004818   NA 36 30 1.013253 -0.2301693
## 4:     %Air       sec 0.04551 0.0004818   NA 36 30 1.013253 -0.2350784
## 5:     %Air       sec 0.04551 0.0004818   NA 36 30 1.013253 -0.2301813
## 6:     %Air       sec 0.04551 0.0004818   NA 36 30 1.013253 -0.2065119
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -505.8623          NA  mgO2/hr/kg   -505.8623
## 2:   -469.1244          NA  mgO2/hr/kg   -469.1244
## 3:   -477.7280          NA  mgO2/hr/kg   -477.7280
## 4:   -487.9171          NA  mgO2/hr/kg   -487.9171
## 5:   -477.7528          NA  mgO2/hr/kg   -477.7528
## 6:   -428.6259          NA  mgO2/hr/kg   -428.6259
## -----------------------------------------
## remove lowest slope 
apoly_rmr <- apoly_rmr |> 
  filter(rate.output != max(rate.output))

Results

results <- data.frame(Clutch = Clutch, 
                      Replicate =Replicate, 
                      Male=Male, 
                      Female=Female,
                      Population = Population, 
                      Tank = Tank,
                      Mass = mass, 
                      Chamber = chamber, 
                      System = system1,
                      Volume = chamber_vol, 
                      Date_tested = Date_tested, 
                      Date_analysed =Date_analysed,
                      Swim = Swim,
                      Salinity = salinity, 
                      Temperature = as.numeric(unique(firesting2$temperature)), 
                      Resting_kg = mean(apoly_rmr$rate.output*-1), 
                      Resting =  mean(apoly_rmr$rate.output*-1)*mass, 
                      rsqrest =mean(apoly_rmr$rsq))
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest
104 6 CPRE447 CPRE452 Pretty patches 171 0.0004818 ch3 Asus 0.04551 2023-05-16 2024-06-14 good/good 36 30 483.6769 0.2330355 0.9846

Maximum oxygen consumption

Data manipulation

firesting2_mmr <- firesting_mmr |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_mmr, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch3
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 1.87
## -----------------------------------------

Subset data

cycle1.start <-  Cycle_1.mmr[1,1]
cycle1.end <-  tail(Cycle_1.mmr, n=1)[1,1] 

cycle1.start.row <- which(firesting2_mmr$TIME == cycle1.start); cycle1.start
## Warning in which(firesting2_mmr$TIME == cycle1.start): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1.end.row <- which(firesting2_mmr$TIME == cycle1.end); cycle1.end 
## Warning in which(firesting2_mmr$TIME == cycle1.end): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1_data <- firesting2_mmr |> 
  subset_data(from = cycle1.start.row, 
              to = cycle1.end.row, 
              by = "row") 
## subset_data: Multi-column dataset detected in input! 
## subset_data is generally intended to subset data already passed through inspect(), or 2-column data frames where time and oxygen are in columns 1 and 2 respectively. 
## Subsetting will proceed anyway based on this assumption, but please ensure you understand what you are doing.
inspect(cycle1_data)
## inspect: Applying column default of 'time = 1'
## inspect: Applying column default of 'oxygen = 2'
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch3
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  9 10 11 12 13 14 16 17 19 20 21 22 23
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 1.51
## -----------------------------------------

Calculating MMR

mmr <- auto_rate(cycle1_data, method = "highest", plot=TRUE, width=60, by="time") |> 
  summary()
## Warning: auto_rate: Multi-column dataset detected in input. Selecting first two columns by default.
##   If these are not the intended data, inspect() or subset the data frame columns appropriately before running auto_rate()

## 
## # summary.auto_rate # -------------------
## 
## === Summary of Results by Highest Rate ===
##      rep rank intercept_b0    slope_b1       rsq density row endrow    time
##   1:  NA    1     615.6827 -0.07293176 0.9916788      NA 119    164 7208.67
##   2:  NA    2     615.4348 -0.07289683 0.9916269      NA 120    165 7210.03
##   3:  NA    3     613.5621 -0.07263940 0.9912478      NA 118    163 7207.31
##   4:  NA    4     613.3335 -0.07260604 0.9912309      NA 121    166 7211.39
##   5:  NA    5     611.0418 -0.07228918 0.9908891      NA 122    167 7212.76
##  ---                                                                       
## 173:  NA  173     273.9057 -0.02547214 0.8731548      NA  71    116 7143.32
## 174:  NA  174     269.5545 -0.02486004 0.8707105      NA  67    112 7137.88
## 175:  NA  175     267.5603 -0.02458645 0.8770803      NA  70    115 7141.96
## 176:  NA  176     263.8517 -0.02406631 0.8830168      NA  68    113 7139.25
## 177:  NA  177     263.3848 -0.02400274 0.8838996      NA  69    114 7140.60
##      endtime    oxy endoxy        rate
##   1: 7268.67 89.860 85.602 -0.07293176
##   2: 7270.03 89.834 85.590 -0.07289683
##   3: 7267.31 89.908 85.642 -0.07263940
##   4: 7271.39 89.812 85.541 -0.07260604
##   5: 7272.76 89.736 85.436 -0.07228918
##  ---                                  
## 173: 7203.32 91.932 90.070 -0.02547214
## 174: 7197.88 92.741 90.651 -0.02486004
## 175: 7201.96 92.094 90.132 -0.02458645
## 176: 7199.25 92.555 90.481 -0.02406631
## 177: 7200.60 92.290 90.321 -0.02400274
## 
## Regressions : 177 | Results : 177 | Method : highest | Roll width : 60 | Roll type : time 
## -----------------------------------------

Adjusting

mmr_adj <- adjust_rate(mmr, by=bg_pre, method = "mean");mmr_adj
## adjust_rate: Rate adjustments applied using "mean" method.
## 
## # print.adjust_rate # -------------------
## NOTE: Consider the sign of the adjustment value when adjusting the rate.
## 
## Adjustment was applied using the 'mean' method.
## 
## Rank 1 of 177 adjusted rate(s):
## Rate          : -0.07293176
## Adjustment    : -0.0003964544
## Adjusted Rate : -0.07253531 
## 
## To see other results use 'pos' input.
## To see full results use summary().
## -----------------------------------------

Converting units

mmr_adj2 <- mmr_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253)
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.

selecting rates

mmr_final <- mmr_adj2 |> 
  select_rate(method = "rsq", n=c(0.93,1)) |> 
  select_rate(method = "highest", n=1) |> 
  plot(type="full") |> 
  summary(export=TRUE)
## select_rate: Selecting rates with rsq values between 0.93 and 1...
## ----- Selection complete. 44 rate(s) removed, 133 rate(s) remaining -----
## select_rate: Selecting highest 1 *absolute* rate values...
## ----- Selection complete. 132 rate(s) removed, 1 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1       rsq density row endrow    time
## 1:  NA    1     615.6827 -0.07293176 0.9916788      NA 119    164 7208.67
##    endtime   oxy endoxy        rate    adjustment rate.adjusted  rate.input
## 1: 7268.67 89.86 85.602 -0.07293176 -0.0003964544   -0.07253531 -0.07253531
##    oxy.unit time.unit  volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.04551 0.0004818   NA 36 30 1.013253 -0.7350132
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -1525.557          NA  mgO2/hr/kg   -1525.557
## -----------------------------------------

Results

results <-  results |> 
  mutate(Max_kg = mmr_final$rate.output*-1, 
         Max = (mmr_final$rate.output*-1)*mass,
         rsqmax =mmr_final$rsq,
         AAS_kg = Max_kg - Resting_kg, 
         AAS = Max - Resting, 
         Notes=Notes, 
         True_resting="") 
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest Max_kg Max rsqmax AAS_kg AAS Notes True_resting
104 6 CPRE447 CPRE452 Pretty patches 171 0.0004818 ch3 Asus 0.04551 2023-05-16 2024-06-14 good/good 36 30 483.6769 0.2330355 0.9846 1525.557 0.7350132 0.9916788 1041.88 0.5019776
### Expor ting data
resp_results_juveniles <- read_csv("resp_results_juveniles.csv") 
## Rows: 98 Columns: 25
## ── Column specification ────────────────────────────────────────────────────────
## Delimiter: ","
## chr  (9): Male, Female, Population, Chamber, System, Date_tested, Swim, Note...
## dbl (16): Clutch, Replicate, Tank, Mass, Volume, Date_analysed, Salinity, Te...
## 
## ℹ Use `spec()` to retrieve the full column specification for this data.
## ℹ Specify the column types or set `show_col_types = FALSE` to quiet this message.
resp_results_juveniles <- rbind(resp_results_juveniles, results) 
resp_results_juveniles 
write.csv(resp_results_juveniles, file="./resp_results_juveniles.csv", row.names = FALSE)

7

Enter specimen data

Replicate = 7 
mass = 0.0005107 
chamber = "ch2" 
Swim = "good/good"
chamber_vol = chamber2_asus
system1 = "Asus"
Notes=""

##--- time of trail ---## 
experiment_mmr_date_asus <- "16 May 2023 12 27PM/Oxygen"
experiment_mmr_date2_asus <- "16 May 2023 12 27PM/All"

firesting_mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",experiment_mmr_date_asus,"data raw/Firesting.txt"), 
    delim = "\t", escape_double = FALSE, 
    col_types = cols(`Time (HH:MM:SS)` = col_time(format = "%H:%M:%S"), 
        `Time (s)` = col_number(), Ch1...5 = col_number(), 
        Ch2...6 = col_number(), Ch3...7 = col_number(), 
        Ch4...8 = col_number()), trim_ws = TRUE, 
    skip = 19) 
## New names:
## • `Ch1` -> `Ch1...5`
## • `Ch2` -> `Ch2...6`
## • `Ch3` -> `Ch3...7`
## • `Ch4` -> `Ch4...8`
## • `Ch 1` -> `Ch 1...9`
## • `Ch 2` -> `Ch 2...10`
## • `Ch 3` -> `Ch 3...11`
## • `Ch 4` -> `Ch 4...12`
## • `('C)` -> `('C)...15`
## • `('C)` -> `('C)...16`
## • `Ch 1` -> `Ch 1...18`
## • `Ch 2` -> `Ch 2...19`
## • `Ch 3` -> `Ch 3...20`
## • `Ch 4` -> `Ch 4...21`
## • `Ch1` -> `Ch1...22`
## • `Ch2` -> `Ch2...23`
## • `Ch3` -> `Ch3...24`
## • `Ch4` -> `Ch4...25`
## • `Ch1` -> `Ch1...26`
## • `Ch2` -> `Ch2...27`
## • `Ch3` -> `Ch3...28`
## • `Ch4` -> `Ch4...29`
## • `` -> `...31`
## Warning: One or more parsing issues, call `problems()` on your data frame for details,
## e.g.:
##   dat <- vroom(...)
##   problems(dat)
Cycle_1.mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",experiment_mmr_date2_asus,"slopes/Cycle_1.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 
## New names:
## • `` -> `...8`

Background rates

Pre-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",preexperiment_date_asus,"slopes")) 

pre_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

pre_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

pre_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))


bg_pre1 <- pre_cycle1 %>% calc_rate.bg()
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre2 <- pre_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre3 <- pre_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre <- mean(bg_pre1$rate.bg.mean,bg_pre2$rate.bg.mean,bg_pre3$rate.bg.mean) 
bg_pre
## [1] 0.0006344115

post-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",postexperiment_date_asus,"slopes")) 
 

post_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

post_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

post_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

bg_post1 <- post_cycle1 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post2 <- post_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post3 <- post_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post <- mean(bg_post1$rate.bg.mean,bg_post2$rate.bg.mean,bg_post3$rate.bg.mean)
bg_post 
## [1] 0.0001945326

Resting metabolic rate

Data manipulation

firesting2_asus <- firesting_asus |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_asus, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch2
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 5.08
## -----------------------------------------

#### subset data

Tstart.row=which(firesting2_asus$TIME ==Cycle_1_asus$Time[1], firesting_asus$TIME) 
Tstart.dTIME=as.numeric(firesting2_asus[Tstart.row, "dTIME"]) 

Tend.row=which(firesting2_asus$TIME ==tail(Cycle_last_asus$Time, n=1), firesting_asus$TIME) 
Tend.dTIME=as.numeric(firesting2_asus[Tend.row, "dTIME"])  

apoly_insp <- firesting2_asus |> 
  subset_data(from=Tstart.dTIME, 
              to=Tend.dTIME, 
              by="time") 

apoly_insp <- inspect(apoly_insp, time=1, oxygen=2)
## 
## # print.inspect # -----------------------
##                 dTIME  ch2
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  4  5  6  7  8  9 10 11 12 13 14 15 16 17 19 20 21 23
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 5.08
## -----------------------------------------

Extract rates

apoly_cr.int <- calc_rate.int(apoly_insp, 
                              starts=(195+45+300), 
                              wait=45, 
                              measure=245, 
                              by="time", 
                              plot=TRUE) 
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from all replicates ...
## plot.calc_rate.int: Plotting first 20 selected reps only. To plot others modify 'pos' input.

## -----------------------------------------

adjust rates for background

apoly_cr.int_adj <- adjust_rate(apoly_cr.int, 
                                by = bg_pre, 
                                by2 = bg_post, 
                                time_by = Tstart.row, 
                                time_by2 = Tend.row,
                                method = "linear")
## Warning: adjust_rate: One or more of the timestamps for the rate(s) in 'x' do not lie between the timestamps for the 'by' and 'by2' background rates. 
## Ensure this is correct. The adjustment value has been calculated regardless by extrapolating outside the background rates time window.
## adjust_rate: Rate adjustments applied using "linear" method.
apoly_cr.int_adj$summary

Converting units

apoly_cr.int_adj2 <- apoly_cr.int_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253) 
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.
apoly_cr.int_adj2$summary

Plot curve

ggplot(as.data.frame(apoly_cr.int_adj2$summary), aes(x=row, y=rate.output*-1)) + 
  geom_point() + 
  stat_smooth(method = "lm", formula = y~poly(x, 2), color="red") +
  theme_classic()

Rate filtering

apoly_rmr <- apoly_cr.int_adj2 |> 
  select_rate(method ="rsq", n=c(0.95,1)) |> 
  select_rate(method="lowest", n=6) |> 
  plot(type="full") |> 
  summary(export = TRUE)
## select_rate: Selecting rates with rsq values between 0.95 and 1...
## ----- Selection complete. 2 rate(s) removed, 19 rate(s) remaining -----
## select_rate: Selecting lowest 6 *absolute* rate values...
## ----- Selection complete. 13 rate(s) removed, 6 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1   rsq density  row endrow     time
## 1:  12    1     343.8552 -0.01710432 0.986      NA 4258   4439 14298.41
## 2:  14    1     407.9710 -0.02001064 0.957      NA 5055   5236 15379.41
## 3:  15    1     396.3746 -0.01863141 0.985      NA 5453   5633 15919.70
## 4:  16    1     411.7717 -0.01894531 0.986      NA 5850   6030 16458.92
## 5:  18    1     421.3196 -0.01833808 0.965      NA 6645   6825 17539.79
## 6:  19    1     402.0031 -0.01672043 0.992      NA 7042   7223 18079.16
##     endtime    oxy endoxy        rate    adjustment rate.adjusted  rate.input
## 1: 14544.15 98.997 95.024 -0.01710432  1.786332e-04   -0.01728296 -0.01728296
## 2: 15625.05 99.576 94.887 -0.02001064  1.194267e-04   -0.02013006 -0.02013006
## 3: 16164.60 99.540 95.195 -0.01863141  8.985391e-05   -0.01872126 -0.01872126
## 4: 16703.61 99.451 95.119 -0.01894531  6.032517e-05   -0.01900564 -0.01900564
## 5: 17784.46 99.131 95.075 -0.01833808  1.123629e-06   -0.01833920 -0.01833920
## 6: 18324.95 99.390 95.620 -0.01672043 -2.844975e-05   -0.01669198 -0.01669198
##    oxy.unit time.unit  volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.04573 0.0005107   NA 36 30 1.013253 -0.1759779
## 2:     %Air       sec 0.04573 0.0005107   NA 36 30 1.013253 -0.2049676
## 3:     %Air       sec 0.04573 0.0005107   NA 36 30 1.013253 -0.1906229
## 4:     %Air       sec 0.04573 0.0005107   NA 36 30 1.013253 -0.1935185
## 5:     %Air       sec 0.04573 0.0005107   NA 36 30 1.013253 -0.1867327
## 6:     %Air       sec 0.04573 0.0005107   NA 36 30 1.013253 -0.1699604
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -344.5817          NA  mgO2/hr/kg   -344.5817
## 2:   -401.3464          NA  mgO2/hr/kg   -401.3464
## 3:   -373.2582          NA  mgO2/hr/kg   -373.2582
## 4:   -378.9279          NA  mgO2/hr/kg   -378.9279
## 5:   -365.6408          NA  mgO2/hr/kg   -365.6408
## 6:   -332.7989          NA  mgO2/hr/kg   -332.7989
## -----------------------------------------
## remove lowest slope 
apoly_rmr <- apoly_rmr |> 
  filter(rate.output != max(rate.output))

Results

results <- data.frame(Clutch = Clutch, 
                      Replicate =Replicate, 
                      Male=Male, 
                      Female=Female,
                      Population = Population, 
                      Tank = Tank,
                      Mass = mass, 
                      Chamber = chamber, 
                      System = system1,
                      Volume = chamber_vol, 
                      Date_tested = Date_tested, 
                      Date_analysed =Date_analysed,
                      Swim = Swim,
                      Salinity = salinity, 
                      Temperature = as.numeric(unique(firesting2$temperature)), 
                      Resting_kg = mean(apoly_rmr$rate.output*-1), 
                      Resting =  mean(apoly_rmr$rate.output*-1)*mass, 
                      rsqrest =mean(apoly_rmr$rsq))
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest
104 7 CPRE447 CPRE452 Pretty patches 171 0.0005107 ch2 Asus 0.04573 2023-05-16 2024-06-14 good/good 36 30 372.751 0.1903639 0.9758

Maximum oxygen consumption

Data manipulation

firesting2_mmr <- firesting_mmr |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_mmr, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch2
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 1.87
## -----------------------------------------

Subset data

cycle1.start <-  Cycle_1.mmr[1,1]
cycle1.end <-  tail(Cycle_1.mmr, n=1)[1,1] 

cycle1.start.row <- which(firesting2_mmr$TIME == cycle1.start); cycle1.start
## Warning in which(firesting2_mmr$TIME == cycle1.start): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1.end.row <- which(firesting2_mmr$TIME == cycle1.end); cycle1.end 
## Warning in which(firesting2_mmr$TIME == cycle1.end): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1_data <- firesting2_mmr |> 
  subset_data(from = cycle1.start.row, 
              to = cycle1.end.row,  
              by = "row") 
## subset_data: Multi-column dataset detected in input! 
## subset_data is generally intended to subset data already passed through inspect(), or 2-column data frames where time and oxygen are in columns 1 and 2 respectively. 
## Subsetting will proceed anyway based on this assumption, but please ensure you understand what you are doing.
inspect(cycle1_data)
## inspect: Applying column default of 'time = 1'
## inspect: Applying column default of 'oxygen = 2'
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch2
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  2  3  4  5  7  8  9 10 11 12 13 14 15 16 17 18 19 20 21 22
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 1.66
## -----------------------------------------

Calculating MMR

mmr <- auto_rate(cycle1_data, method = "highest", plot=TRUE, width=60, by="time") |> 
  summary()
## Warning: auto_rate: Multi-column dataset detected in input. Selecting first two columns by default.
##   If these are not the intended data, inspect() or subset the data frame columns appropriately before running auto_rate()

## 
## # summary.auto_rate # -------------------
## 
## === Summary of Results by Highest Rate ===
##      rep rank intercept_b0    slope_b1       rsq density row endrow    time
##   1:  NA    1     671.9694 -0.07498145 0.9948532      NA   4     49 7649.24
##   2:  NA    2     671.8659 -0.07496894 0.9948357      NA   3     48 7647.91
##   3:  NA    3     670.8713 -0.07484040 0.9945655      NA   2     47 7646.55
##   4:  NA    4     670.2263 -0.07475372 0.9944657      NA   5     50 7650.60
##   5:  NA    5     669.2096 -0.07462489 0.9941252      NA   1     46 7645.16
##  ---                                                                       
## 172:  NA  172     376.4364 -0.03679887 0.9939835      NA 115    160 7800.44
## 173:  NA  173     374.9013 -0.03660062 0.9961453      NA 111    156 7795.00
## 174:  NA  174     374.4566 -0.03654519 0.9956794      NA 114    159 7799.09
## 175:  NA  175     373.5539 -0.03642865 0.9964192      NA 112    157 7796.37
## 176:  NA  176     373.1467 -0.03637716 0.9966530      NA 113    158 7797.74
##      endtime    oxy endoxy        rate
##   1: 7709.24 98.341 94.082 -0.07498145
##   2: 7707.91 98.399 94.156 -0.07496894
##   3: 7706.55 98.477 94.227 -0.07484040
##   4: 7710.60 98.336 94.009 -0.07475372
##   5: 7705.16 98.545 94.298 -0.07462489
##  ---                                  
## 172: 7860.44 89.518 86.985 -0.03679887
## 173: 7855.00 89.741 87.445 -0.03660062
## 174: 7859.09 89.565 87.088 -0.03654519
## 175: 7856.37 89.663 87.358 -0.03642865
## 176: 7857.74 89.638 87.223 -0.03637716
## 
## Regressions : 176 | Results : 176 | Method : highest | Roll width : 60 | Roll type : time 
## -----------------------------------------

Adjusting

mmr_adj <- adjust_rate(mmr, by=bg_pre, method = "mean");mmr_adj
## adjust_rate: Rate adjustments applied using "mean" method.
## 
## # print.adjust_rate # -------------------
## NOTE: Consider the sign of the adjustment value when adjusting the rate.
## 
## Adjustment was applied using the 'mean' method.
## 
## Rank 1 of 176 adjusted rate(s):
## Rate          : -0.07498145
## Adjustment    : 0.0006344115
## Adjusted Rate : -0.07561586 
## 
## To see other results use 'pos' input.
## To see full results use summary().
## -----------------------------------------

Converting units

mmr_adj2 <- mmr_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253)
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.

selecting rates

mmr_final <- mmr_adj2 |> 
  select_rate(method = "rsq", n=c(0.93,1)) |> 
  select_rate(method = "highest", n=1) |> 
  plot(type="full") |> 
  summary(export=TRUE)
## select_rate: Selecting rates with rsq values between 0.93 and 1...
## ----- Selection complete. 0 rate(s) removed, 176 rate(s) remaining -----
## select_rate: Selecting highest 1 *absolute* rate values...
## ----- Selection complete. 175 rate(s) removed, 1 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1       rsq density row endrow    time
## 1:  NA    1     671.9694 -0.07498145 0.9948532      NA   4     49 7649.24
##    endtime    oxy endoxy        rate   adjustment rate.adjusted  rate.input
## 1: 7709.24 98.341 94.082 -0.07498145 0.0006344115   -0.07561586 -0.07561586
##    oxy.unit time.unit  volume      mass area  S  t        P  rate.abs
## 1:     %Air       sec 0.04573 0.0005107   NA 36 30 1.013253 -0.769933
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -1507.603          NA  mgO2/hr/kg   -1507.603
## -----------------------------------------

Results

results <-  results |> 
  mutate(Max_kg = mmr_final$rate.output*-1, 
         Max = (mmr_final$rate.output*-1)*mass,
         rsqmax =mmr_final$rsq,
         AAS_kg = Max_kg - Resting_kg, 
         AAS = Max - Resting, 
         Notes=Notes, 
         True_resting="") 
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest Max_kg Max rsqmax AAS_kg AAS Notes True_resting
104 7 CPRE447 CPRE452 Pretty patches 171 0.0005107 ch2 Asus 0.04573 2023-05-16 2024-06-14 good/good 36 30 372.751 0.1903639 0.9758 1507.603 0.769933 0.9948532 1134.852 0.5795691
### Expor ting data
resp_results_juveniles <- read_csv("resp_results_juveniles.csv") 
## Rows: 99 Columns: 25
## ── Column specification ────────────────────────────────────────────────────────
## Delimiter: ","
## chr  (9): Male, Female, Population, Chamber, System, Date_tested, Swim, Note...
## dbl (16): Clutch, Replicate, Tank, Mass, Volume, Date_analysed, Salinity, Te...
## 
## ℹ Use `spec()` to retrieve the full column specification for this data.
## ℹ Specify the column types or set `show_col_types = FALSE` to quiet this message.
resp_results_juveniles <- rbind(resp_results_juveniles, results) 
resp_results_juveniles 
write.csv(resp_results_juveniles, file="./resp_results_juveniles.csv", row.names = FALSE)

8

Enter specimen data

Replicate = 8 
mass = 0.0006159 
chamber = "ch1" 
Swim = "good/good"
chamber_vol = chamber1_asus
system1 = "Asus"
Notes=""

##--- time of trail ---## 
experiment_mmr_date_asus <- "16 May 2023 12 38PM/Oxygen"
experiment_mmr_date2_asus <- "16 May 2023 12 38PM/All"

firesting_mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",experiment_mmr_date_asus,"data raw/Firesting.txt"), 
    delim = "\t", escape_double = FALSE, 
    col_types = cols(`Time (HH:MM:SS)` = col_time(format = "%H:%M:%S"), 
        `Time (s)` = col_number(), Ch1...5 = col_number(), 
        Ch2...6 = col_number(), Ch3...7 = col_number(), 
        Ch4...8 = col_number()), trim_ws = TRUE, 
    skip = 19) 
## New names:
## • `Ch1` -> `Ch1...5`
## • `Ch2` -> `Ch2...6`
## • `Ch3` -> `Ch3...7`
## • `Ch4` -> `Ch4...8`
## • `Ch 1` -> `Ch 1...9`
## • `Ch 2` -> `Ch 2...10`
## • `Ch 3` -> `Ch 3...11`
## • `Ch 4` -> `Ch 4...12`
## • `('C)` -> `('C)...15`
## • `('C)` -> `('C)...16`
## • `Ch 1` -> `Ch 1...18`
## • `Ch 2` -> `Ch 2...19`
## • `Ch 3` -> `Ch 3...20`
## • `Ch 4` -> `Ch 4...21`
## • `Ch1` -> `Ch1...22`
## • `Ch2` -> `Ch2...23`
## • `Ch3` -> `Ch3...24`
## • `Ch4` -> `Ch4...25`
## • `Ch1` -> `Ch1...26`
## • `Ch2` -> `Ch2...27`
## • `Ch3` -> `Ch3...28`
## • `Ch4` -> `Ch4...29`
## • `` -> `...31`
## Warning: One or more parsing issues, call `problems()` on your data frame for details,
## e.g.:
##   dat <- vroom(...)
##   problems(dat)
Cycle_1.mmr <- read_delim(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",experiment_mmr_date2_asus,"slopes/Cycle_1.txt"), 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        `Seconds from start for linreg` = col_number(), 
        `ch1 po2` = col_number(), `ch2 po2` = col_number(), 
        `ch3 po2` = col_number(), `ch4 po2` = col_number(), 
        ...8 = col_skip()), trim_ws = TRUE) 
## New names:
## • `` -> `...8`

Background rates

Pre-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",preexperiment_date_asus,"slopes")) 

pre_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

pre_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

pre_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))


bg_pre1 <- pre_cycle1 %>% calc_rate.bg()
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre2 <- pre_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre3 <- pre_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_pre <- mean(bg_pre1$rate.bg.mean,bg_pre2$rate.bg.mean,bg_pre3$rate.bg.mean) 
bg_pre
## [1] -0.0004832052

post-experiment

setwd(paste("C:/Users/jc527762/OneDrive - James Cook University/PhD dissertation/Data/2023/Resp_backup/2023_Resp/Asus/Experiment_",postexperiment_date_asus,"slopes")) 
 

post_cycle1 <- read_delim("./Cycle_1.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

post_cycle2 <- read_delim("./Cycle_2.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber)) 

post_cycle3 <- read_delim("./Cycle_3.txt", 
    delim = ";", escape_double = FALSE, col_types = cols(Time = col_time(format = "%H:%M:%S"), 
        ...8 = col_skip()), trim_ws = TRUE) %>% 
  rename(dTIME = `Seconds from start for linreg`, 
         ch1 =`ch1 po2`, 
         ch2 =`ch2 po2`, 
         ch3 =`ch3 po2`, 
         ch4 =`ch4 po2`) %>% 
  select(c("Time",chamber))

bg_post1 <- post_cycle1 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post2 <- post_cycle2 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post3 <- post_cycle3 %>% calc_rate.bg() 
## 
## # plot.calc_rate.bg # -------------------

## plot.calc_rate.bg: Plotting all 1 background rates ...
## -----------------------------------------
bg_post <- mean(bg_post1$rate.bg.mean,bg_post2$rate.bg.mean,bg_post3$rate.bg.mean)
bg_post 
## [1] -0.001557929

Resting metabolic rate

Data manipulation

firesting2_asus <- firesting_asus |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_asus, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch1
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 5.08
## -----------------------------------------

#### subset data

Tstart.row=which(firesting2_asus$TIME ==Cycle_1_asus$Time[1], firesting_asus$TIME) 
Tstart.dTIME=as.numeric(firesting2_asus[Tstart.row, "dTIME"]) 

Tend.row=which(firesting2_asus$TIME ==tail(Cycle_last_asus$Time, n=1), firesting_asus$TIME) 
Tend.dTIME=as.numeric(firesting2_asus[Tend.row, "dTIME"])  

apoly_insp <- firesting2_asus |> 
  subset_data(from=Tstart.dTIME, 
              to=Tend.dTIME, 
              by="time") 

apoly_insp <- inspect(apoly_insp, time=1, oxygen=2)
## 
## # print.inspect # -----------------------
##                 dTIME  ch1
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  4  5  6  7  8  9 10 11 12 13 14 15 16 17 19 20 21 23
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 5.08
## -----------------------------------------

Extract rates

apoly_cr.int <- calc_rate.int(apoly_insp, 
                              starts=(195+45+300), 
                              wait=45, 
                              measure=245, 
                              by="time", 
                              plot=TRUE) 
## 
## # plot.calc_rate.int # ------------------
## plot.calc_rate.int: Plotting rate from all replicates ...
## plot.calc_rate.int: Plotting first 20 selected reps only. To plot others modify 'pos' input.

## -----------------------------------------

adjust rates for background

apoly_cr.int_adj <- adjust_rate(apoly_cr.int, 
                                by = bg_pre, 
                                by2 = bg_post, 
                                time_by = Tstart.row, 
                                time_by2 = Tend.row,
                                method = "linear")
## Warning: adjust_rate: One or more of the timestamps for the rate(s) in 'x' do not lie between the timestamps for the 'by' and 'by2' background rates. 
## Ensure this is correct. The adjustment value has been calculated regardless by extrapolating outside the background rates time window.
## adjust_rate: Rate adjustments applied using "linear" method.
apoly_cr.int_adj$summary

Converting units

apoly_cr.int_adj2 <- apoly_cr.int_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253) 
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.
apoly_cr.int_adj2$summary

Plot curve

ggplot(as.data.frame(apoly_cr.int_adj2$summary), aes(x=row, y=rate.output*-1)) + 
  geom_point() + 
  stat_smooth(method = "lm", formula = y~poly(x, 2), color="red") +
  theme_classic()

Rate filtering

apoly_rmr <- apoly_cr.int_adj2 |> 
  select_rate(method ="rsq", n=c(0.95,1)) |> 
  select_rate(method="lowest", n=6) |> 
  plot(type="full") |> 
  summary(export = TRUE)
## select_rate: Selecting rates with rsq values between 0.95 and 1...
## ----- Selection complete. 0 rate(s) removed, 21 rate(s) remaining -----
## select_rate: Selecting lowest 6 *absolute* rate values...
## ----- Selection complete. 15 rate(s) removed, 6 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1   rsq density  row endrow     time
## 1:   6    1     350.1719 -0.02286707 0.988      NA 2018   2198 11058.76
## 2:   7    1     345.1105 -0.02134123 0.984      NA 2415   2596 11598.75
## 3:   8    1     362.5785 -0.02185788 0.995      NA 2813   2993 12138.70
## 4:  10    1     403.6346 -0.02316393 0.991      NA 3608   3770 13219.12
## 5:  11    1     415.8305 -0.02314164 0.989      NA 3904   4041 13759.09
## 6:  14    1     472.0083 -0.02434495 0.994      NA 5055   5236 15379.41
##     endtime    oxy endoxy        rate   adjustment rate.adjusted  rate.input
## 1: 11303.55 96.888 91.195 -0.02286707 -0.001163175   -0.02170389 -0.02170389
## 2: 11844.57 97.008 92.077 -0.02134123 -0.001235507   -0.02010572 -0.02010572
## 3: 12383.32 96.974 91.830 -0.02185788 -0.001307684   -0.02055020 -0.02055020
## 4: 13463.66 97.089 91.600 -0.02316393 -0.001452262   -0.02171167 -0.02171167
## 5: 14003.80 96.988 91.416 -0.02314164 -0.001524534   -0.02161711 -0.02161711
## 6: 15625.05 97.302 91.662 -0.02434495 -0.001741430   -0.02260352 -0.02260352
##    oxy.unit time.unit  volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.04565 0.0006159   NA 36 30 1.013253 -0.2206060
## 2:     %Air       sec 0.04565 0.0006159   NA 36 30 1.013253 -0.2043616
## 3:     %Air       sec 0.04565 0.0006159   NA 36 30 1.013253 -0.2088794
## 4:     %Air       sec 0.04565 0.0006159   NA 36 30 1.013253 -0.2206850
## 5:     %Air       sec 0.04565 0.0006159   NA 36 30 1.013253 -0.2197238
## 6:     %Air       sec 0.04565 0.0006159   NA 36 30 1.013253 -0.2297500
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -358.1847          NA  mgO2/hr/kg   -358.1847
## 2:   -331.8097          NA  mgO2/hr/kg   -331.8097
## 3:   -339.1450          NA  mgO2/hr/kg   -339.1450
## 4:   -358.3130          NA  mgO2/hr/kg   -358.3130
## 5:   -356.7525          NA  mgO2/hr/kg   -356.7525
## 6:   -373.0314          NA  mgO2/hr/kg   -373.0314
## -----------------------------------------
## remove lowest slope 
apoly_rmr <- apoly_rmr |> 
  filter(rate.output != max(rate.output))

Results

results <- data.frame(Clutch = Clutch, 
                      Replicate =Replicate, 
                      Male=Male, 
                      Female=Female,
                      Population = Population, 
                      Tank = Tank,
                      Mass = mass, 
                      Chamber = chamber, 
                      System = system1,
                      Volume = chamber_vol, 
                      Date_tested = Date_tested, 
                      Date_analysed =Date_analysed,
                      Swim = Swim,
                      Salinity = salinity, 
                      Temperature = as.numeric(unique(firesting2$temperature)), 
                      Resting_kg = mean(apoly_rmr$rate.output*-1), 
                      Resting =  mean(apoly_rmr$rate.output*-1)*mass, 
                      rsqrest =mean(apoly_rmr$rsq))
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest
104 8 CPRE447 CPRE452 Pretty patches 171 0.0006159 ch1 Asus 0.04565 2023-05-16 2024-06-14 good/good 36 30 357.0853 0.2199289 0.9914

Maximum oxygen consumption

Data manipulation

firesting2_mmr <- firesting_mmr |>
  select(c(1:3,5:9)) |> 
  rename(TIME = `Time (HH:MM:SS)`, 
         dTIME = `Time (s)`, 
         ch1 = Ch1...5, 
         ch2 = Ch2...6,
         ch3 = Ch3...7, 
         ch4 = Ch4...8, 
         temperature= `Ch 1...9`) |> 
  select(c("dTIME",all_of(chamber),"TIME","temperature"))

Inspect file

inspect(firesting2_mmr, time=1, oxygen=2)
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch1
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  3  4  5  6  7  8  9 10 11 12 13 14 15 16 17 18 19 20
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 5.08
## -----------------------------------------

Subset data

cycle1.start <-  Cycle_1.mmr[1,1]
cycle1.end <-  tail(Cycle_1.mmr, n=1)[1,1] 

cycle1.start.row <- which(firesting2_mmr$TIME == cycle1.start); cycle1.start
## Warning in which(firesting2_mmr$TIME == cycle1.start): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1.end.row <- which(firesting2_mmr$TIME == cycle1.end); cycle1.end 
## Warning in which(firesting2_mmr$TIME == cycle1.end): Incompatible methods
## ("Ops.difftime", "Ops.data.frame") for "=="
cycle1_data <- firesting2_mmr |> 
  subset_data(from = cycle1.start.row, 
              to = cycle1.end.row, 
              by = "row") 
## subset_data: Multi-column dataset detected in input! 
## subset_data is generally intended to subset data already passed through inspect(), or 2-column data frames where time and oxygen are in columns 1 and 2 respectively. 
## Subsetting will proceed anyway based on this assumption, but please ensure you understand what you are doing.
inspect(cycle1_data)
## inspect: Applying column default of 'time = 1'
## inspect: Applying column default of 'oxygen = 2'
## Warning: inspect: Time values are not evenly-spaced (numerically).
## inspect: Data issues detected. For more information use print().
## 
## # print.inspect # -----------------------
##                 dTIME  ch1
## numeric          pass pass
## Inf/-Inf         pass pass
## NA/NaN           pass pass
## sequential       pass    -
## duplicated       pass    -
## evenly-spaced    WARN    -
## 
## Uneven Time data locations (first 20 shown) in column: dTIME 
##  [1]  1  2  4  5  6  7  8  9 10 11 12 13 14 15 16 17 19 20 21 23
## Minimum and Maximum intervals in uneven Time data: 
## [1] 1.33 1.65
## -----------------------------------------

Calculating MMR

mmr <- auto_rate(cycle1_data, method = "highest", plot=TRUE, width=60, by="time") |> 
  summary()
## Warning: auto_rate: Multi-column dataset detected in input. Selecting first two columns by default.
##   If these are not the intended data, inspect() or subset the data frame columns appropriately before running auto_rate()

## 
## # summary.auto_rate # -------------------
## 
## === Summary of Results by Highest Rate ===
##      rep rank intercept_b0    slope_b1       rsq density row endrow    time
##   1:  NA    1     659.9519 -0.06743896 0.9858514      NA  44     89 8373.91
##   2:  NA    2     659.2415 -0.06735532 0.9856429      NA  43     88 8372.55
##   3:  NA    3     658.2431 -0.06723451 0.9851489      NA  45     90 8375.27
##   4:  NA    4     655.9414 -0.06696328 0.9847268      NA  42     87 8371.19
##   5:  NA    5     654.8158 -0.06682568 0.9837589      NA  46     91 8376.66
##  ---                                                                       
## 172:  NA  172     210.6901 -0.01415131 0.9504998      NA  87    132 8432.70
## 173:  NA  173     210.1806 -0.01409124 0.9495781      NA  86    131 8431.35
## 174:  NA  174     210.1791 -0.01409079 0.9496662      NA  88    133 8434.06
## 175:  NA  175     210.1174 -0.01408370 0.9497470      NA  89    134 8435.43
## 176:  NA  176     209.2952 -0.01398676 0.9477664      NA  85    130 8429.97
##      endtime    oxy endoxy        rate
##   1: 8433.91 95.122 91.326 -0.06743896
##   2: 8432.55 95.159 91.318 -0.06735532
##   3: 8435.27 95.037 91.356 -0.06723451
##   4: 8431.19 95.172 91.317 -0.06696328
##   5: 8436.66 94.999 91.322 -0.06682568
##  ---                                  
## 172: 8492.70 91.317 90.520 -0.01415131
## 173: 8491.35 91.531 90.537 -0.01409124
## 174: 8494.06 91.318 90.529 -0.01409079
## 175: 8495.43 91.326 90.437 -0.01408370
## 176: 8489.97 91.564 90.510 -0.01398676
## 
## Regressions : 176 | Results : 176 | Method : highest | Roll width : 60 | Roll type : time 
## -----------------------------------------

Adjusting

mmr_adj <- adjust_rate(mmr, by=bg_pre, method = "mean");mmr_adj
## adjust_rate: Rate adjustments applied using "mean" method.
## 
## # print.adjust_rate # -------------------
## NOTE: Consider the sign of the adjustment value when adjusting the rate.
## 
## Adjustment was applied using the 'mean' method.
## 
## Rank 1 of 176 adjusted rate(s):
## Rate          : -0.06743896
## Adjustment    : -0.0004832052
## Adjusted Rate : -0.06695575 
## 
## To see other results use 'pos' input.
## To see full results use summary().
## -----------------------------------------

Converting units

mmr_adj2 <- mmr_adj |> 
  convert_rate(oxy.unit = "%Air", 
               time.unit = "secs", 
               output.unit = "mg/h/kg", 
               volume = chamber_vol,
               mass = mass,
               S = salinity, 
               t = as.numeric(unique(firesting2$temperature)), 
               P = 1.013253)
## convert_rate: Object of class 'adjust_rate' detected. Converting all adjusted rates in '$rate.adjusted'.

selecting rates

mmr_final <- mmr_adj2 |> 
  select_rate(method = "rsq", n=c(0.93,1)) |> 
  select_rate(method = "highest", n=1) |> 
  plot(type="full") |> 
  summary(export=TRUE)
## select_rate: Selecting rates with rsq values between 0.93 and 1...
## ----- Selection complete. 32 rate(s) removed, 144 rate(s) remaining -----
## select_rate: Selecting highest 1 *absolute* rate values...
## ----- Selection complete. 143 rate(s) removed, 1 rate(s) remaining -----
## 
## # plot.convert_rate # -------------------
## plot.convert_rate: Plotting all rate(s)...

## -----------------------------------------
## 
## # summary.convert_rate # ----------------
## Summary of all converted rates:
## 
##    rep rank intercept_b0    slope_b1       rsq density row endrow    time
## 1:  NA    1     659.9519 -0.06743896 0.9858514      NA  44     89 8373.91
##    endtime    oxy endoxy        rate    adjustment rate.adjusted  rate.input
## 1: 8433.91 95.122 91.326 -0.06743896 -0.0004832052   -0.06695575 -0.06695575
##    oxy.unit time.unit  volume      mass area  S  t        P   rate.abs
## 1:     %Air       sec 0.04565 0.0006159   NA 36 30 1.013253 -0.6805617
##    rate.m.spec rate.a.spec output.unit rate.output
## 1:   -1104.987          NA  mgO2/hr/kg   -1104.987
## -----------------------------------------

Results

results <-  results |> 
  mutate(Max_kg = mmr_final$rate.output*-1, 
         Max = (mmr_final$rate.output*-1)*mass,
         rsqmax =mmr_final$rsq,
         AAS_kg = Max_kg - Resting_kg, 
         AAS = Max - Resting, 
         Notes=Notes, 
         True_resting="") 
knitr::kable(results, "simple") 
Clutch Replicate Male Female Population Tank Mass Chamber System Volume Date_tested Date_analysed Swim Salinity Temperature Resting_kg Resting rsqrest Max_kg Max rsqmax AAS_kg AAS Notes True_resting
104 8 CPRE447 CPRE452 Pretty patches 171 0.0006159 ch1 Asus 0.04565 2023-05-16 2024-06-14 good/good 36 30 357.0853 0.2199289 0.9914 1104.987 0.6805617 0.9858514 747.9021 0.4606329
### Expor ting data
resp_results_juveniles <- read_csv("resp_results_juveniles.csv") 
## Rows: 100 Columns: 25
## ── Column specification ────────────────────────────────────────────────────────
## Delimiter: ","
## chr  (9): Male, Female, Population, Chamber, System, Date_tested, Swim, Note...
## dbl (16): Clutch, Replicate, Tank, Mass, Volume, Date_analysed, Salinity, Te...
## 
## ℹ Use `spec()` to retrieve the full column specification for this data.
## ℹ Specify the column types or set `show_col_types = FALSE` to quiet this message.
resp_results_juveniles <- rbind(resp_results_juveniles, results) 
resp_results_juveniles 
write.csv(resp_results_juveniles, file="./resp_results_juveniles.csv", row.names = FALSE)